Generation of knock-in primary human T cells using Cas9 ribonucleoproteins

Schumann, Kathrin; Lin, Steven; Boyer, Eric; Simeonov, Dimitre R.; Subramaniam, Meena; Gate, Rachel E.; Haliburton, Genevieve E.; Ye, Chun J.; Bluestone, Jeffrey A.; Doudna, Jennifer A.; Marson, Alexander

doi:10.1073/pnas.1512503112

Title: Generation of knock-in primary human T cells using Cas9 ribonucleoproteins

Journal Article · Mon Jul 27 00:00:00 EDT 2015 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1512503112· OSTI ID:1221821

Schumann, Kathrin ^[1]; Lin, Steven ^[2]; Boyer, Eric ^[1]; Simeonov, Dimitre R. ^[1]; Subramaniam, Meena ^[1]; Gate, Rachel E. ^[1]; Haliburton, Genevieve E. ^[1]; Ye, Chun J. ^[1]; Bluestone, Jeffrey A. ^[1]; Doudna, Jennifer A. ^[3]; Marson, Alexander ^[4]

Univ. of California, San Francisco, CA (United States)
Univ. of California, Berkeley, CA (United States)
Univ. of California, San Francisco, CA (United States); Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Univ. of California, San Francisco, CA (United States); Univ. of California, Berkeley, CA (United States)

T-cell genome engineering holds great promise for cell-based therapies for cancer, HIV, primary immune deficiencies, and autoimmune diseases, but genetic manipulation of human T cells has been challenging. Improved tools are needed to efficiently “knock out” genes and “knock in” targeted genome modifications to modulate T-cell function and correct disease-associated mutations. CRISPR/Cas9 technology is facilitating genome engineering in many cell types, but in human T cells its efficiency has been limited and it has not yet proven useful for targeted nucleotide replacements. Here we report efficient genome engineering in human CD4⁺ T cells using Cas9:single-guide RNA ribonucleoproteins (Cas9 RNPs). Cas9 RNPs allowed ablation of CXCR4, a coreceptor for HIV entry. Cas9 RNP electroporation caused up to ~40% of cells to lose high-level cell-surface expression of CXCR4, and edited cells could be enriched by sorting based on low CXCR4 expression. Importantly, Cas9 RNPs paired with homology-directed repair template oligonucleotides generated a high frequency of targeted genome modifications in primary T cells. Targeted nucleotide replacement was achieved in CXCR4 and PD-1 (PDCD1), a regulator of T-cell exhaustion that is a validated target for tumor immunotherapy. Deep sequencing of a target site confirmed that Cas9 RNPs generated knock-in genome modifications with up to ~20% efficiency, which accounted for up to approximately one-third of total editing events. These results establish Cas9 RNP technology for diverse experimental and therapeutic genome engineering applications in primary human T cells.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1221821

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 112, Issue 33; ISSN 0027-8424

Publisher:: National Academy of Sciences, Washington, DC (United States)Copyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 456 works

Citation information provided by
Web of Science

References (24)

Adoptive Immunotherapy for Cancer or Viruses Maus, Marcela V.; Fraietta, Joseph A.; Levine, Bruce L. Annual Review of Immunology, Vol. 32, Issue 1 https://doi.org/10.1146/annurev-immunol-032713-120136	journal	March 2014
Helper T-cell identity and evolution of differential transcriptomes and epigenomes Vahedi, Golnaz; C. Poholek, Amanda; Hand, Timothy W. Immunological Reviews, Vol. 252, Issue 1 https://doi.org/10.1111/imr.12037	journal	February 2013
HIV-1 Entry Cofactor: Functional cDNA Cloning of a Seven-Transmembrane, G Protein-Coupled Receptor Feng, Y.; Broder, C. C.; Kennedy, P. E. Science, Vol. 272, Issue 5263 https://doi.org/10.1126/science.272.5263.872	journal	May 1996
Chimeric Antigen Receptor–Modified T Cells in Chronic Lymphoid Leukemia Porter, David L.; Levine, Bruce L.; Kalos, Michael New England Journal of Medicine, Vol. 365, Issue 8 https://doi.org/10.1056/NEJMoa1103849	journal	August 2011
Adoptive immunotherapy for cancer: harnessing the T cell response Restifo, Nicholas P.; Dudley, Mark E.; Rosenberg, Steven A. Nature Reviews Immunology, Vol. 12, Issue 4 https://doi.org/10.1038/nri3191	journal	March 2012
Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9 Mandal, Pankaj K.; Ferreira, Leonardo M. R.; Collins, Ryan Cell Stem Cell, Vol. 15, Issue 5 https://doi.org/10.1016/j.stem.2014.10.004	journal	November 2014
Multifactorial T-cell Hypofunction That Is Reversible Can Limit the Efficacy of Chimeric Antigen Receptor-Transduced Human T cells in Solid Tumors Moon, E. K.; Wang, L. -C.; Dolfi, D. V. Clinical Cancer Research, Vol. 20, Issue 16 https://doi.org/10.1158/1078-0432.CCR-13-2627	journal	June 2014
Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases Sung, Y. H.; Kim, J. M.; Kim, H. -T. Genome Research, Vol. 24, Issue 1 https://doi.org/10.1101/gr.163394.113	journal	November 2013
Targeted genome editing in human repopulating haematopoietic stem cells Genovese, Pietro; Schiroli, Giulia; Escobar, Giulia Nature, Vol. 510, Issue 7504 https://doi.org/10.1038/nature13420	journal	May 2014
Gene Editing of CCR5 in Autologous CD4 T Cells of Persons Infected with HIV Tebas, Pablo; Stein, David; Tang, Winson W. New England Journal of Medicine, Vol. 370, Issue 10 https://doi.org/10.1056/NEJMoa1300662	journal	March 2014
Long-Term Control of HIV by CCR5 Delta32/Delta32 Stem-Cell Transplantation Hütter, Gero; Nowak, Daniel; Mossner, Maximilian New England Journal of Medicine, Vol. 360, Issue 7 https://doi.org/10.1056/NEJMoa0802905	journal	February 2009
Genetic and epigenetic fine mapping of causal autoimmune disease variants Farh, Kyle Kai-How; Marson, Alexander; Zhu, Jiang Nature, Vol. 518, Issue 7539 https://doi.org/10.1038/nature13835	journal	October 2014
CD4+ T Cells from IPEX Patients Convert into Functional and Stable Regulatory T Cells by FOXP3 Gene Transfer Passerini, L.; Mel, E. R.; Sartirana, C. Science Translational Medicine, Vol. 5, Issue 215 https://doi.org/10.1126/scitranslmed.3007320	journal	December 2013
Anti-PD-1 Antibody Therapy Potently Enhances the Eradication of Established Tumors By Gene-Modified T Cells John, L. B.; Devaud, C.; Duong, C. P. M. Clinical Cancer Research, Vol. 19, Issue 20 https://doi.org/10.1158/1078-0432.CCR-13-0458	journal	July 2013
The new frontier of genome engineering with CRISPR-Cas9 Doudna, Jennifer A.; Charpentier, Emmanuelle Science, Vol. 346, Issue 6213 https://doi.org/10.1126/science.1258096	journal	November 2014
Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery Lin, Steven; Staahl, Brett T.; Alla, Ravi K. eLife, Vol. 3 https://doi.org/10.7554/eLife.04766	journal	December 2014
Immune Checkpoint Blockade: A Common Denominator Approach to Cancer Therapy Topalian, Suzanne L.; Drake, Charles G.; Pardoll, Drew M. Cancer Cell, Vol. 27, Issue 4 https://doi.org/10.1016/j.ccell.2015.03.001	journal	April 2015
Development and Applications of CRISPR-Cas9 for Genome Engineering Hsu, Patrick D.; Lander, Eric S.; Zhang, Feng Cell, Vol. 157, Issue 6 https://doi.org/10.1016/j.cell.2014.05.010	journal	June 2014
Function of the chemokine receptor CXCR4 in haematopoiesis and in cerebellar development Zou, Yong-Rui; Kottmann, Andreas H.; Kuroda, Masahiko Nature, Vol. 393, Issue 6685 https://doi.org/10.1038/31269	journal	June 1998
Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins Kim, S.; Kim, D.; Cho, S. W. Genome Research, Vol. 24, Issue 6 https://doi.org/10.1101/gr.171322.113	journal	April 2014
Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo Zuris, John A.; Thompson, David B.; Shu, Yilai Nature Biotechnology, Vol. 33, Issue 1 https://doi.org/10.1038/nbt.3081	journal	October 2014
Simultaneous zinc-finger nuclease editing of the HIV coreceptors ccr5 and cxcr4 protects CD4+ T cells from HIV-1 infection Didigu, Chuka A.; Wilen, Craig B.; Wang, Jianbin Blood, Vol. 123, Issue 1 https://doi.org/10.1182/blood-2013-08-521229	journal	January 2014
Double-Strand Break End Resection and Repair Pathway Choice Symington, Lorraine S.; Gautier, Jean Annual Review of Genetics, Vol. 45, Issue 1 https://doi.org/10.1146/annurev-genet-110410-132435	journal	December 2011
A seven-transmembrane domain receptor involved in fusion and entry of T-cell-tropic human immunodeficiency virus type 1 strains. Berson, J. F.; Long, D.; Doranz, B. J. Journal of virology, Vol. 70, Issue 9 https://doi.org/10.1128/jvi.70.9.6288-6295.1996	journal	January 1996

Cited By (166)

Acoustophoretic rapid media exchange and continuous-flow electrotransfection of primary human T cells for applications in automated cellular therapy manufacturing Hsi, Peter; Christianson, Rebecca J.; Dubay, Ryan A. Lab on a Chip, Vol. 19, Issue 18 https://doi.org/10.1039/c9lc00458k	journal	January 2019
The next generation of CRISPR–Cas technologies and applications Pickar-Oliver, Adrian; Gersbach, Charles A. Nature Reviews Molecular Cell Biology, Vol. 20, Issue 8 https://doi.org/10.1038/s41580-019-0131-5	journal	May 2019
Efficient precise knockin with a double cut HDR donor after CRISPR/Cas9-mediated double-stranded DNA cleavage Zhang, Jian-Ping; Li, Xiao-Lan; Li, Guo-Hua Genome Biology, Vol. 18, Issue 1 https://doi.org/10.1186/s13059-017-1164-8	journal	February 2017
Genome editing by natural and engineered CRISPR-associated nucleases Wu, Wen Y.; Lebbink, Joyce H. G.; Kanaar, Roland Nature Chemical Biology, Vol. 14, Issue 7 https://doi.org/10.1038/s41589-018-0080-x	journal	June 2018
CRISPR/Cas9 genome editing in human hematopoietic stem cells Bak, Rasmus O.; Dever, Daniel P.; Porteus, Matthew H. Nature Protocols, Vol. 13, Issue 2 https://doi.org/10.1038/nprot.2017.143	journal	January 2018
Delivery of CRISPR/Cas9 by Novel Strategies for Gene Therapy Wang, Le; Zheng, Wenfu; Liu, Shaoqin ChemBioChem https://doi.org/10.1002/cbic.201800629	journal	December 2018
Genome editing of bread wheat using biolistic delivery of CRISPR/Cas9 in vitro transcripts or ribonucleoproteins Liang, Zhen; Chen, Kunling; Zhang, Yi Nature Protocols, Vol. 13, Issue 3 https://doi.org/10.1038/nprot.2017.145	journal	February 2018
Advancing chimeric antigen receptor T cell therapy with CRISPR/Cas9 Ren, Jiangtao; Zhao, Yangbing Protein & Cell, Vol. 8, Issue 9 https://doi.org/10.1007/s13238-017-0410-x	journal	April 2017
Orthotopic replacement of T-cell receptor α- and β-chains with preservation of near-physiological T-cell function Schober, Kilian; Müller, Thomas R.; Gökmen, Füsun Nature Biomedical Engineering, Vol. 3, Issue 12 https://doi.org/10.1038/s41551-019-0409-0	journal	June 2019
Helios enhances the preferential differentiation of human fetal CD4 ⁺ naïve T cells into regulatory T cells Ng, Melissa S. F.; Roth, Theodore L.; Mendoza, Ventura F. Science Immunology, Vol. 4, Issue 41 https://doi.org/10.1126/sciimmunol.aav5947	journal	November 2019
Application of Genome Editing Techniques in Immunology Zych, Agata O.; Bajor, Malgorzata; Zagozdzon, Radoslaw Archivum Immunologiae et Therapiae Experimentalis, Vol. 66, Issue 4 https://doi.org/10.1007/s00005-018-0504-z	journal	January 2018
Therapeutic applications of CRISPR RNA-guided genome editing Koo, Taeyoung; Kim, Jin-Soo Briefings in Functional Genomics, Vol. 16, Issue 1 https://doi.org/10.1093/bfgp/elw032	journal	August 2016
Strategies for nonviral nanoparticle‐based delivery of CRISPR/Cas9 therapeutics Chen, Fengqian; Alphonse, Martin; Liu, Qi WIREs Nanomedicine and Nanobiotechnology https://doi.org/10.1002/wnan.1609	journal	December 2019
The human leukemia virus HTLV-1 alters the structure and transcription of host chromatin in cis Melamed, Anat; Yaguchi, Hiroko; Miura, Michi eLife https://doi.org/10.1101/277335	journal	March 2018
CRISPR/Cas9-Mediated Genome Editing in Epstein-Barr Virus-Transformed Lymphoblastoid B-Cell Lines: CRISPR in EBV-Transformed B-Cells Jiang, Sizun; Wang, Liang Wei; Walsh, Michael J. Current Protocols in Molecular Biology, Vol. 121, Issue 1 https://doi.org/10.1002/cpmb.51	journal	January 2018
Emerging CRISPR/Cas9 applications for T-cell gene editing Preece, Roland; Georgiadis, Christos Emerging Topics in Life Sciences, Vol. 3, Issue 3 https://doi.org/10.1042/ETLS20180144	journal	April 2019
CRISPR/Cas9 Genome Editing Using Gold-Nanoparticle-Mediated Laserporation Bošnjak, Berislav; Permanyer, Marc; Sethi, Maya K. Advanced Biosystems, Vol. 2, Issue 11 https://doi.org/10.1002/adbi.201700184	journal	May 2018
Metabolic reprogramming of human CD8 ⁺ memory T cells through loss of SIRT1 Jeng, Mark Y.; Hull, Philip A.; Fei, Mingjian The Journal of Experimental Medicine, Vol. 215, Issue 1 https://doi.org/10.1084/jem.20161066	journal	November 2017
Functional Interrogation of Primary Human T Cells via CRISPR Genetic Editing Chen, Xin; Kozhaya, Lina; Tastan, Cihan The Journal of Immunology, Vol. 201, Issue 5 https://doi.org/10.4049/jimmunol.1701616	journal	July 2018
Delivery technologies for genome editing Yin, Hao; Kauffman, Kevin J.; Anderson, Daniel G. Nature Reviews Drug Discovery, Vol. 16, Issue 6 https://doi.org/10.1038/nrd.2016.280	journal	March 2017
An efficient electroporation protocol for the genetic modification of mammalian cells Chicaybam, Leonardo; Barcelos, Camila; Peixoto, Barbara Frontiers in Bioengineering and Biotechnology https://doi.org/10.1101/073387	journal	September 2016
CRISPR-delivery particles targeting nuclear receptor–interacting protein 1 ( Nrip1 ) in adipose cells to enhance energy expenditure Shen, Yuefei; Cohen, Jessica L.; Nicoloro, Sarah M. Journal of Biological Chemistry, Vol. 293, Issue 44 https://doi.org/10.1074/jbc.ra118.004554	journal	September 2018
Transfection by Electroporation Potter, Huntington; Heller, Richard Current Protocols in Molecular Biology, Vol. 121, Issue 1 https://doi.org/10.1002/cpmb.48	journal	January 2018
Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing Yin, Hao; Song, Chun-Qing; Suresh, Sneha Nature Biotechnology, Vol. 35, Issue 12 https://doi.org/10.1038/nbt.4005	journal	November 2017
Recent advances in DNA-free editing and precise base editing in plants Zhang, Yi; Gao, Caixia Emerging Topics in Life Sciences, Vol. 1, Issue 2 https://doi.org/10.1042/etls20170021	journal	November 2017
Genome-wide CRISPR Screens in Primary Human T Cells Reveal Key Regulators of Immune Function Shifrut, Eric; Carnevale, Julia; Tobin, Victoria Cell, Vol. 175, Issue 7 https://doi.org/10.1016/j.cell.2018.10.024	journal	December 2018
Delivering CRISPR: a review of the challenges and approaches Lino, Christopher A.; Harper, Jason C.; Carney, James P. Drug Delivery, Vol. 25, Issue 1 https://doi.org/10.1080/10717544.2018.1474964	journal	January 2018
Highly efficient and versatile plasmid-based gene editing in primary T cells Kornete, Mara; Marone, Romina; Jeker, Lukas T. The Journal of Immunology https://doi.org/10.1101/247544	posted_content	January 2018
Resources for the design of CRISPR gene editing experiments Graham, Daniel B.; Root, David E. Genome Biology, Vol. 16, Issue 1 https://doi.org/10.1186/s13059-015-0823-x	journal	November 2015
Viable and efficient electroporation-based genetic manipulation of unstimulated human T cells Aksoy, Pinar; Aksoy, Bülent Arman; Czech, Eric BioRxiv https://doi.org/10.1101/466243	posted_content	October 2019
Cancer diagnosis and immunotherapy in the age of CRISPR Cook, Peter J.; Ventura, Andrea Genes, Chromosomes and Cancer, Vol. 58, Issue 4 https://doi.org/10.1002/gcc.22702	journal	December 2018
CRISPR/Cas9: A tool for immunological research Hochheiser, Katharina; Kueh, Andrew J.; Gebhardt, Thomas European Journal of Immunology, Vol. 48, Issue 4 https://doi.org/10.1002/eji.201747131	journal	February 2018
Recent challenges and advances in genetically-engineered cell therapy Yong, Seok-Beom; Chung, Jee Young; Song, Yoonsung Journal of Pharmaceutical Investigation, Vol. 48, Issue 2 https://doi.org/10.1007/s40005-017-0381-1	journal	December 2017
CRISPR/Cas9 technology as a potent molecular tool for gene therapy Karimian, Ansar; Azizian, Khalil; Parsian, Hadi Journal of Cellular Physiology, Vol. 234, Issue 8 https://doi.org/10.1002/jcp.27972	journal	December 2018
An efficient and scalable pipeline for epitope tagging in mammalian stem cells using Cas9 ribonucleoprotein Dewari, Pooran Singh; Southgate, Benjamin; Mccarten, Katrina eLife https://doi.org/10.1101/255737	journal	January 2018
CRISPR/Cas-mediated knock-in via non-homologous end-joining in the crustacean Daphnia magna Kumagai, Hitoshi; Nakanishi, Takashi; Matsuura, Tomoaki PLOS ONE, Vol. 12, Issue 10 https://doi.org/10.1371/journal.pone.0186112	journal	October 2017
CRISPR–Cas9 genome engineering of primary CD4+ T cells for the interrogation of HIV–host factor interactions Hultquist, Judd F.; Hiatt, Joseph; Schumann, Kathrin Nature Protocols, Vol. 14, Issue 1 https://doi.org/10.1038/s41596-018-0069-7	journal	December 2018
HIV-1 inhibition in cells with CXCR4 mutant genome created by CRISPR-Cas9 and piggyBac recombinant technologies Liu, Shuai; Wang, Qiankun; Yu, Xiao Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-018-26894-4	journal	June 2018
Engineered T cells: the promise and challenges of cancer immunotherapy Fesnak, Andrew D.; June, Carl H.; Levine, Bruce L. Nature Reviews Cancer, Vol. 16, Issue 9 https://doi.org/10.1038/nrc.2016.97	journal	August 2016
CRISPR-Cas9-mediated multiplex gene editing in CAR-T cells Liu, Xiaojuan; Zhang, Yongping; Cheng, Chen Cell Research, Vol. 27, Issue 1 https://doi.org/10.1038/cr.2016.142	journal	December 2016
Resolving mechanisms of immune-mediated disease in primary CD4 T cells. Bourges, Christophe; Groff, Abigail F.; Burren, Oliver Apollo - University of Cambridge Repository https://doi.org/10.17863/cam.51450	text	January 2020
In vitro–transcribed guide RNAs trigger an innate immune response via the RIG-I pathway Wienert, Beeke; Shin, Jiyung; Zelin, Elena PLOS Biology, Vol. 16, Issue 7 https://doi.org/10.1371/journal.pbio.2005840	journal	July 2018
Applications of CRISPR systems in respiratory health: Entering a new ‘red pen’ era in genome editing Moses, Colette; Kaur, Parwinder Respirology, Vol. 24, Issue 7 https://doi.org/10.1111/resp.13527	journal	February 2019
Genome-wide CRISPR Screens in Primary Human T Cells Reveal Key Regulators of Immune Function Shifrut, Eric; Carnevale, Julia; Tobin, Victoria Cell https://doi.org/10.1101/384776	journal	August 2018
Primary allogeneic mitochondrial mix (PAMM) transfer/transplant by MitoCeption to address damage in PBMCs caused by ultraviolet radiation Cabrera, Francisco; Ortega, Mayra; Velarde, Francesca BMC Biotechnology, Vol. 19, Issue 1 https://doi.org/10.1186/s12896-019-0534-6	journal	June 2019
The clinical potential of gene editing as a tool to engineer cell‐based therapeutics Ashmore‐Harris, Candice; Fruhwirth, Gilbert O. Clinical and Translational Medicine, Vol. 9, Issue 1 https://doi.org/10.1186/s40169-020-0268-z	journal	January 2020
Genetic abrogation of immune checkpoints in antigen-specific cytotoxic T-lymphocyte as a potential alternative to blockade immunotherapy Zhang, Chi; Peng, Yanchun; Hublitz, Philip Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-018-23803-7	journal	April 2018
Carboxylated branched poly(β-amino ester) nanoparticles enable robust cytosolic protein delivery and CRISPR-Cas9 gene editing Rui, Yuan; Wilson, David R.; Choi, John Science Advances, Vol. 5, Issue 12 https://doi.org/10.1126/sciadv.aay3255	journal	December 2019
Selection-free genome editing of the sickle mutation in human adult hematopoietic stem/progenitor cells DeWitt, M. A.; Magis, W.; Bray, N. L. Science Translational Medicine, Vol. 8, Issue 360 https://doi.org/10.1126/scitranslmed.aaf9336	journal	October 2016
GapmeR cellular internalization by macropinocytosis induces sequence-specific gene silencing in human primary T-cells Fazil, Mobashar Hussain Urf Turabe; Ong, Seow Theng; Chalasani, Madhavi Latha Somaraju Scientific Reports, Vol. 6, Issue 1 https://doi.org/10.1038/srep37721	journal	November 2016
Targeted gene editing restores regulated CD40L function in X-linked hyper-IgM syndrome Hubbard, Nicholas; Hagin, David; Sommer, Karen Blood, Vol. 127, Issue 21 https://doi.org/10.1182/blood-2015-11-683235	journal	May 2016
Epigenetic changes around the pX region and spontaneous HTLV-1 transcription are CTCF-independent Miura, Michi; Miyazato, Paola; Satou, Yorifumi Wellcome Open Research, Vol. 3 https://doi.org/10.12688/wellcomeopenres.14741.2	journal	January 2018
Spontaneous HTLV-1 transcription is accompanied by distinct epigenetic changes in the 5′ and 3′ long terminal repeats Miura, Michi; Miyazato, Paola; Satou, Yorifumi Wellcome Open Research, Vol. 3 https://doi.org/10.12688/wellcomeopenres.14741.1	journal	January 2018
Application of CRISPR/Cas9-Based Gene Editing in HIV-1/AIDS Therapy Xiao, Qiaoqiao; Guo, Deyin; Chen, Shuliang Frontiers in Cellular and Infection Microbiology, Vol. 9 https://doi.org/10.3389/fcimb.2019.00069	journal	March 2019
Efficient CRISPR/Cas9 Disruption of Autoimmune-Associated Genes Reveals Key Signaling Programs in Primary Human T Cells Anderson, Warren; Thorpe, Jerill; Long, S. Alice The Journal of Immunology, Vol. 203, Issue 12 https://doi.org/10.4049/jimmunol.1900848	journal	November 2019
Highly Efficient and Versatile Plasmid-Based Gene Editing in Primary T Cells Kornete, Mara; Marone, Romina; Jeker, Lukas T. The Journal of Immunology, Vol. 200, Issue 7 https://doi.org/10.4049/jimmunol.1701121	journal	February 2018
Lessons learned from the blockade of immune checkpoints in cancer immunotherapy Li, Xiaolei; Shao, Changshun; Shi, Yufang Journal of Hematology & Oncology, Vol. 11, Issue 1 https://doi.org/10.1186/s13045-018-0578-4	journal	February 2018
Minimal 2'-O-methyl phosphorothioate linkage modification pattern of synthetic guide RNAs for increased stability and efficient CRISPR-Cas9 gene editing avoiding cellular toxicity Basila, Megan; Kelley, Melissa L.; Smith, Anja van Brabant PLOS ONE, Vol. 12, Issue 11 https://doi.org/10.1371/journal.pone.0188593	journal	November 2017
Genetic editing and interrogation with Cpf1 and caged truncated pre-tRNA-like crRNA in mammalian cells Zhang, Xuhua; Xu, Linping; Fan, Ruihua Cell Discovery, Vol. 4, Issue 1 https://doi.org/10.1038/s41421-018-0035-0	journal	July 2018
Purity of transferred CD8 ⁺ T cells is crucial for safety and efficacy of combinatorial tumor immunotherapy in the absence of SHP‐1 Watson, H. Angharad; Dolton, Garry; Ohme, Julia Immunology & Cell Biology, Vol. 94, Issue 8 https://doi.org/10.1038/icb.2016.45	journal	July 2016
Extension of the crRNA enhances Cpf1 gene editing in vitro and in vivo Park, Hyo Min; Liu, Hui; Wu, Joann Nature Communications, Vol. 9, Issue 1 https://doi.org/10.1038/s41467-018-05641-3	journal	August 2018
Allele-specific expression changes dynamically during T cell activation in HLA and other autoimmune loci Gutierrez-Arcelus, Maria; Baglaenko, Yuriy; Arora, Jatin Nature Genetics, Vol. 52, Issue 3 https://doi.org/10.1038/s41588-020-0579-4	journal	February 2020
Electroporation of mice zygotes with dual guide RNA/Cas9 complexes for simple and efficient cloning-free genome editing Teixeira, Marie; Py, Bénédicte F.; Bosc, Christophe Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-017-18826-5	journal	January 2018
An Efficient Electroporation Protocol for the Genetic Modification of Mammalian Cells Chicaybam, Leonardo; Barcelos, Camila; Peixoto, Barbara Frontiers in Bioengineering and Biotechnology, Vol. 4 https://doi.org/10.3389/fbioe.2016.00099	journal	January 2017
Ribonucleoprotein Transfection for CRISPR/Cas9-Mediated Gene Knockout in Primary T Cells Oh, Soyoung A.; Seki, Akiko; Rutz, Sascha Current Protocols in Immunology, Vol. 124, Issue 1 https://doi.org/10.1002/cpim.69	journal	October 2018
CRISPR/Cas9-mediated PD-1 disruption enhances anti-tumor efficacy of human chimeric antigen receptor T cells Rupp, Levi J.; Schumann, Kathrin; Roybal, Kole T. Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/s41598-017-00462-8	journal	April 2017
Efficient gene knockin in axolotl and its use to test the role of satellite cells in limb regeneration Fei, Ji-Feng; Schuez, Maritta; Knapp, Dunja Proceedings of the National Academy of Sciences, Vol. 114, Issue 47 https://doi.org/10.1073/pnas.1706855114	journal	October 2017
CRISPR/Cas9 system targeting regulatory genes of HIV-1 inhibits viral replication in infected T-cell cultures Ophinni, Youdiil; Inoue, Mari; Kotaki, Tomohiro Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-018-26190-1	journal	May 2018
Therapeutic potential of CRISPR/Cas9 gene editing in engineered T‐cell therapy Gao, Qianqian; Dong, Xuan; Xu, Qumiao Cancer Medicine, Vol. 8, Issue 9 https://doi.org/10.1002/cam4.2257	journal	June 2019
Directed evolution of CRISPR-Cas9 to increase its specificity Lee, Jungjoon K.; Jeong, Euihwan; Lee, Joonsun Nature Communications, Vol. 9, Issue 1 https://doi.org/10.1038/s41467-018-05477-x	journal	August 2018
Gene Editing of HIV-1 Co-receptors to Prevent and/or Cure Virus Infection Allen, Alexander G.; Chung, Cheng-Han; Atkins, Andrew Frontiers in Microbiology, Vol. 9 https://doi.org/10.3389/fmicb.2018.02940	journal	December 2018
Cornerstones of CRISPR–Cas in drug discovery and therapy Fellmann, Christof; Gowen, Benjamin G.; Lin, Pei-Chun Nature Reviews Drug Discovery, Vol. 16, Issue 2 https://doi.org/10.1038/nrd.2016.238	journal	December 2016
Efficient and versatile CRISPR engineering of human neurons in culture to model neurological disorders Shah, Ruth R.; Cholewa-Waclaw, Justyna; Davies, Faith C. J. Wellcome Open Research, Vol. 1 https://doi.org/10.12688/wellcomeopenres.10011.1	journal	January 2016
CRISPR/Cas9-mediated PD-1 disruption enhances human mesothelin-targeted CAR T cell effector functions Hu, Wanghong; Zi, Zhenguo; Jin, Yanling Cancer Immunology, Immunotherapy, Vol. 68, Issue 3 https://doi.org/10.1007/s00262-018-2281-2	journal	December 2018
Highly efficient genome editing for single-base substitutions using optimized ssODNs with Cas9-RNPs Okamoto, Sachiko; Amaishi, Yasunori; Maki, Izumi Scientific Reports, Vol. 9, Issue 1 https://doi.org/10.1038/s41598-019-41121-4	journal	March 2019
Reprogramming human T cell function and specificity with non-viral genome targeting Roth, Theodore L.; Puig-Saus, Cristina; Yu, Ruby Nature, Vol. 559, Issue 7714 https://doi.org/10.1038/s41586-018-0326-5	journal	July 2018
Human genetic variation alters CRISPR-Cas9 on- and off-targeting specificity at therapeutically implicated loci Lessard, Samuel; Francioli, Laurent; Alfoldi, Jessica Proceedings of the National Academy of Sciences, Vol. 114, Issue 52 https://doi.org/10.1073/pnas.1714640114	journal	December 2017
Light-activated cell identification and sorting (LACIS) for selection of edited clones on a nanofluidic device Mocciaro, Annamaria; Roth, Theodore L.; Bennett, Hayley M. Communications Biology, Vol. 1, Issue 1 https://doi.org/10.1038/s42003-018-0034-6	journal	May 2018
Transmembrane E3 ligase RNF183 mediates ER stress-induced apoptosis by degrading Bcl-xL Wu, Yanfang; Li, Xia; Jia, Junying Proceedings of the National Academy of Sciences, Vol. 115, Issue 12 https://doi.org/10.1073/pnas.1716439115	journal	March 2018
Unexpected heterogeneity derived from Cas9 ribonucleoprotein-introduced clonal cells at the HPRT1 locus Sakuma, Tetsushi; Mochida, Keiji; Nakade, Shota Genes to Cells, Vol. 23, Issue 4 https://doi.org/10.1111/gtc.12569	journal	February 2018
Dynamics of T cell activation mediated by eIF3 interactions with T cell receptor mRNAs De Silva, Dasmanthie; Ferguson, Lucas; Smith, Benjamin E. BioRxiv https://doi.org/10.1101/2019.12.20.885558	posted_content	October 2020
Cas9 Ribonucleoprotein Complex Delivery: Methods and Applications for Neuroinflammation Campbell, Lee A.; Richie, Christopher T.; Maggirwar, Nishad S. Journal of Neuroimmune Pharmacology, Vol. 14, Issue 4 https://doi.org/10.1007/s11481-019-09856-z	journal	June 2019
mRNA as novel technology for passive immunotherapy Schlake, Thomas; Thess, Andreas; Thran, Moritz Cellular and Molecular Life Sciences, Vol. 76, Issue 2 https://doi.org/10.1007/s00018-018-2935-4	journal	October 2018
Nanoparticle delivery of Cas9 ribonucleoprotein and donor DNA in vivo induces homology-directed DNA repair Lee, Kunwoo; Conboy, Michael; Park, Hyo Min Nature Biomedical Engineering, Vol. 1, Issue 11 https://doi.org/10.1038/s41551-017-0137-2	journal	October 2017
A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors Park, Ryan J.; Wang, Tim; Koundakjian, Dylan Nature Genetics, Vol. 49, Issue 2 https://doi.org/10.1038/ng.3741	journal	December 2016
Methods for Enhancing Clustered Regularly Interspaced Short Palindromic Repeats/Cas9-Mediated Homology-Directed Repair Efficiency Tang, Xi-Dian; Gao, Fei; Liu, Ming-Jie Frontiers in Genetics, Vol. 10 https://doi.org/10.3389/fgene.2019.00551	journal	June 2019
Efficient genome editing of human natural killer cells by CRISPR RNP Rautela, Jai; Surgenor, Elliot; Huntington, Nicholas D. BioRxiv https://doi.org/10.1101/406934	posted_content	September 2018
The Potential Use of the CRISPR-Cas System for HIV-1 Gene Therapy Sanches-da-Silva, Gabriela De Nardi; Medeiros, Luiza Fonseca Sales; Lima, Fabio Mitsuo International Journal of Genomics, Vol. 2019 https://doi.org/10.1155/2019/8458263	journal	August 2019
In vitro transcribed guide RNAs trigger an innate immune response via the RIG-I pathway Wienert, Beeke; Shin, Jiyung; Zelin, Elena PLOS Biology https://doi.org/10.1101/275669	journal	March 2018
Editing the Central Nervous System Through CRISPR/Cas9 Systems Cota-Coronado, Agustin; Díaz-Martínez, Néstor Fabián; Padilla-Camberos, Eduardo Frontiers in Molecular Neuroscience, Vol. 12 https://doi.org/10.3389/fnmol.2019.00110	journal	May 2019
Regulatory T cells use arginase 2 to enhance their metabolic fitness in tissues Lowe, Margaret M.; Boothby, Ian; Clancy, Sean JCI Insight, Vol. 4, Issue 24 https://doi.org/10.1172/jci.insight.129756	journal	December 2019
Glucocorticoids paradoxically facilitate steroid resistance in T cell acute lymphoblastic leukemias and thymocytes Meyer, Lauren K.; Huang, Benjamin J.; Delgado-Martin, Cristina Journal of Clinical Investigation, Vol. 130, Issue 2 https://doi.org/10.1172/jci130189	journal	December 2019
A novel method to generate T-cell receptor–deficient chimeric antigen receptor T cells Kamiya, Takahiro; Wong, Desmond; Png, Yi Tian Blood Advances, Vol. 2, Issue 5 https://doi.org/10.1182/bloodadvances.2017012823	journal	March 2018
Guide Swap enables genome-scale pooled CRISPR–Cas9 screening in human primary cells Ting, Pamela Y.; Parker, Albert E.; Lee, J. Scott Nature Methods, Vol. 15, Issue 11 https://doi.org/10.1038/s41592-018-0149-1	journal	October 2018
A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells Vakulskas, Christopher A.; Dever, Daniel P.; Rettig, Garrett R. Nature Medicine, Vol. 24, Issue 8 https://doi.org/10.1038/s41591-018-0137-0	journal	August 2018
Hit-and-run programming of therapeutic cytoreagents using mRNA nanocarriers Moffett, H. F.; Coon, M. E.; Radtke, S. Nature Communications, Vol. 8, Issue 1 https://doi.org/10.1038/s41467-017-00505-8	journal	August 2017
The protean world of non-coding RNAs in glioblastoma Paulmurugan, Ramasamy; Malhotra, Meenakshi; Massoud, Tarik F. Journal of Molecular Medicine, Vol. 97, Issue 7 https://doi.org/10.1007/s00109-019-01798-6	journal	May 2019
Optimized RNP transfection for highly efficient CRISPR/Cas9-mediated gene knockout in primary T cells Seki, Akiko; Rutz, Sascha Journal of Experimental Medicine, Vol. 215, Issue 3 https://doi.org/10.1084/jem.20171626	journal	February 2018
Dysregulated RASGRP1 expression through RUNX1 mediated transcription promotes autoimmunity Baars, Matthijs; Douma, Thera; Simeonov, Dimitre R. European Journal of Immunology https://doi.org/10.1101/656454	journal	November 2019
CRISPR-Cas9 technology: applications and human disease modelling Torres-Ruiz, Raul; Rodriguez-Perales, Sandra Briefings in Functional Genomics, Vol. 16, Issue 1 https://doi.org/10.1093/bfgp/elw025	journal	June 2016
CRISPR/Cas9-based genome editing in the era of CAR T cell immunotherapy Salas-Mckee, January; Kong, Weimin; Gladney, Whitney L. Human Vaccines & Immunotherapeutics, Vol. 15, Issue 5 https://doi.org/10.1080/21645515.2019.1571893	journal	March 2019
In vitro and ex vivo strategies for intracellular delivery Stewart, Martin P.; Sharei, Armon; Ding, Xiaoyun Nature, Vol. 538, Issue 7624 https://doi.org/10.1038/nature19764	journal	October 2016
Genome surgery using Cas9 ribonucleoproteins for the treatment of age-related macular degeneration Kim, Kyoungmi; Park, Sung Wook; Kim, Jin Hyoung Genome Research, Vol. 27, Issue 3 https://doi.org/10.1101/gr.219089.116	journal	February 2017
CRISPR-Cas9: A cornerstone for the evolution of precision medicine Razzouk, Sleiman Annals of Human Genetics, Vol. 82, Issue 6 https://doi.org/10.1111/ahg.12271	journal	July 2018
CRISPR/Cas9 system: A promising technology for the treatment of inherited and neoplastic hematological diseases Antony, Justin S.; Haque, A. K. M. Ashiqul; Lamsfus-Calle, Andrés Advances in Cell and Gene Therapy, Vol. 1, Issue 1 https://doi.org/10.1002/acg2.10	journal	May 2018
Creating and evaluating accurate CRISPR-Cas9 scalpels for genomic surgery Bolukbasi, Mehmet Fatih; Gupta, Ankit; Wolfe, Scot A. Nature Methods, Vol. 13, Issue 1 https://doi.org/10.1038/nmeth.3684	journal	December 2015
Gene editing enables T-cell engineering to redirect antigen specificity for potent tumor rejection Albers, Julian J.; Ammon, Tim; Gosmann, Dario Life Science Alliance, Vol. 2, Issue 2 https://doi.org/10.26508/lsa.201900367	journal	March 2019
Allele-specific expression changes dynamically during T cell activation in HLA and other autoimmune loci Gutierrez-Arcelus, Maria; Baglaenko, Yuriy; Arora, Jatin Nature Genetics https://doi.org/10.1101/599449	journal	April 2019
Next Generation CAR T Cells for the Immunotherapy of High-Grade Glioma Petersen, Christopher T.; Krenciute, Giedre Frontiers in Oncology, Vol. 9 https://doi.org/10.3389/fonc.2019.00069	journal	February 2019
The epigenetic landscape of T cell exhaustion Sen, D. R.; Kaminski, J.; Barnitz, R. A. Science, Vol. 354, Issue 6316 https://doi.org/10.1126/science.aae0491	journal	October 2016
Humanized model mice by genome editing and engraftment technologies Kim, Yong-Sam; Ko, Jeong-Heon Molecular & Cellular Toxicology, Vol. 14, Issue 3 https://doi.org/10.1007/s13273-018-0028-y	journal	June 2018
Is autoimmunity the Achilles' heel of cancer immunotherapy? June, Carl H.; Warshauer, Jeremy T.; Bluestone, Jeffrey A. Nature Medicine, Vol. 23, Issue 5 https://doi.org/10.1038/nm.4321	journal	May 2017
T-Cell Exhaustion in Chronic Infections: Reversing the State of Exhaustion and Reinvigorating Optimal Protective Immune Responses Saeidi, Alireza; Zandi, Keivan; Cheok, Yi Ying Frontiers in Immunology, Vol. 9 https://doi.org/10.3389/fimmu.2018.02569	journal	November 2018
Resolving mechanisms of immune‐mediated disease in primary CD 4 T cells Bourges, Christophe; Groff, Abigail F.; Burren, Oliver S. EMBO Molecular Medicine, Vol. 12, Issue 5 https://doi.org/10.15252/emmm.202012112	journal	April 2020
One-step generation of modular CAR-T cells with AAV–Cpf1 Dai, Xiaoyun; Park, Jonathan J.; Du, Yaying Nature Methods, Vol. 16, Issue 3 https://doi.org/10.1038/s41592-019-0329-7	journal	February 2019
Generation of tumor antigen-specific murine CD8+ T cells with enhanced anti-tumor activity via highly efficient CRISPR/Cas9 genome editing Ouchi, Yasuo; Patil, Ashwini; Tamura, Yusuke International Immunology, Vol. 30, Issue 4 https://doi.org/10.1093/intimm/dxy006	journal	February 2018
Chimeric antigen receptor T cell therapy for multiple myeloma Hasegawa, Kana; Hosen, Naoki Inflammation and Regeneration, Vol. 39, Issue 1 https://doi.org/10.1186/s41232-019-0100-6	journal	June 2019
The implication of CRISPR/Cas9 genome editing technology in combating human oncoviruses Gilani, Usman; Shaukat, Memoona; Rasheed, Arisha Journal of Medical Virology, Vol. 91, Issue 1 https://doi.org/10.1002/jmv.25292	journal	September 2018
CRISPR–Cas: a tool for cancer research and therapeutics Yin, Hao; Xue, Wen; Anderson, Daniel G. Nature Reviews Clinical Oncology, Vol. 16, Issue 5 https://doi.org/10.1038/s41571-019-0166-8	journal	January 2019
Next-generation regulatory T cell therapy Ferreira, Leonardo M. R.; Muller, Yannick D.; Bluestone, Jeffrey A. Nature Reviews Drug Discovery, Vol. 18, Issue 10 https://doi.org/10.1038/s41573-019-0041-4	journal	September 2019
Applications of CRISPR technologies in research and beyond Barrangou, Rodolphe; Doudna, Jennifer A. Nature Biotechnology, Vol. 34, Issue 9 https://doi.org/10.1038/nbt.3659	journal	September 2016
An efficient and scalable pipeline for epitope tagging in mammalian stem cells using Cas9 ribonucleoprotein Dewari, Pooran Singh; Southgate, Benjamin; Mccarten, Katrina eLife, Vol. 7 https://doi.org/10.7554/elife.35069	journal	April 2018
Lipofection-mediated genome editing using DNA-free delivery of the Cas9/gRNA ribonucleoprotein into plant cells Liu, Wusheng; Rudis, Mary R.; Cheplick, Matthew H. Plant Cell Reports, Vol. 39, Issue 2 https://doi.org/10.1007/s00299-019-02488-w	journal	November 2019
EfficientCRISPR‐based genome editing using tandem guideRNAs and editable surrogate reporters Liu, Wuqing; Li, Shifeng; Zhang, Yunbin FEBS Open Bio, Vol. 8, Issue 7 https://doi.org/10.1002/2211-5463.12437	journal	June 2018
In vitro transcribed sgRNA causes cell death by inducing interferon release Mu, Wei; Tang, Na; Cheng, Chen Protein & Cell, Vol. 10, Issue 6 https://doi.org/10.1007/s13238-018-0605-9	journal	January 2019
Stem cell-based therapies for HIV/AIDS Pernet, Olivier; Yadav, Swati Seth; An, Dong Sung Advanced Drug Delivery Reviews, Vol. 103 https://doi.org/10.1016/j.addr.2016.04.027	journal	August 2016
A Cas9 Ribonucleoprotein Platform for Functional Genetic Studies of HIV-Host Interactions in Primary Human T Cells Hultquist, Judd F.; Schumann, Kathrin; Woo, Jonathan M. Cell Reports, Vol. 17, Issue 5 https://doi.org/10.1016/j.celrep.2016.09.080	journal	October 2016
CRISPR/Cas9: a powerful tool for identification of new targets for cancer treatment Liu, Bin; Saber, Ali; Haisma, Hidde J. Drug Discovery Today, Vol. 24, Issue 4 https://doi.org/10.1016/j.drudis.2019.02.011	journal	April 2019
CRISPR-mediated genome editing and human diseases Cai, Liquan; Fisher, Alfred L.; Huang, Haochu Genes & Diseases, Vol. 3, Issue 4 https://doi.org/10.1016/j.gendis.2016.07.003	journal	December 2016
CRISPR-Mediated Editing of the B Cell Receptor in Primary Human B Cells Greiner, Vera; Bou Puerto, Regina; Liu, Suying iScience, Vol. 12 https://doi.org/10.1016/j.isci.2019.01.032	journal	February 2019
Versatility of chemically synthesized guide RNAs for CRISPR-Cas9 genome editing Kelley, Melissa L.; Strezoska, Žaklina; He, Kaizhang Journal of Biotechnology, Vol. 233 https://doi.org/10.1016/j.jbiotec.2016.06.011	journal	September 2016
Delivery of Cas9 Protein into Mouse Zygotes through a Series of Electroporation Dramatically Increases the Efficiency of Model Creation Wang, Wenbo; Kutny, Peter M.; Byers, Shannon L. Journal of Genetics and Genomics, Vol. 43, Issue 5 https://doi.org/10.1016/j.jgg.2016.02.004	journal	May 2016
CRISPR/Cas9: Transcending the Reality of Genome Editing Chira, Sergiu; Gulei, Diana; Hajitou, Amin Molecular Therapy - Nucleic Acids, Vol. 7 https://doi.org/10.1016/j.omtn.2017.04.001	journal	June 2017
CRISPR/Cas9-Directed Reassignment of the GATA1 Initiation Codon in K562 Cells to Recapitulate AML in Down Syndrome Bloh, Kevin M.; Bialk, Pawel A.; Gopalakrishnapillai, Anilkumar Molecular Therapy - Nucleic Acids, Vol. 7 https://doi.org/10.1016/j.omtn.2017.04.009	journal	June 2017
VSV-G-Enveloped Vesicles for Traceless Delivery of CRISPR-Cas9 Montagna, Claudia; Petris, Gianluca; Casini, Antonio Molecular Therapy - Nucleic Acids, Vol. 12 https://doi.org/10.1016/j.omtn.2018.05.010	journal	September 2018
Transient Retrovirus-Based CRISPR/Cas9 All-in-One Particles for Efficient, Targeted Gene Knockout Knopp, Yvonne; Geis, Franziska K.; Heckl, Dirk Molecular Therapy - Nucleic Acids, Vol. 13 https://doi.org/10.1016/j.omtn.2018.09.006	journal	December 2018
Efficient Gene Editing at Major CFTR Mutation Loci Ruan, Jinxue; Hirai, Hiroyuki; Yang, Dongshan Molecular Therapy - Nucleic Acids, Vol. 16 https://doi.org/10.1016/j.omtn.2019.02.006	journal	June 2019
Simplified CRISPR tools for efficient genome editing and streamlined protocols for their delivery into mammalian cells and mouse zygotes Jacobi, Ashley M.; Rettig, Garrett R.; Turk, Rolf Methods, Vol. 121-122 https://doi.org/10.1016/j.ymeth.2017.03.021	journal	May 2017
Integration of a CD19 CAR into the TCR Alpha Chain Locus Streamlines Production of Allogeneic Gene-Edited CAR T Cells MacLeod, Daniel T.; Antony, Jeyaraj; Martin, Aaron J. Molecular Therapy, Vol. 25, Issue 4 https://doi.org/10.1016/j.ymthe.2017.02.005	journal	April 2017
Genome-editing Technologies for Gene and Cell Therapy Maeder, Morgan L.; Gersbach, Charles A. Molecular Therapy, Vol. 24, Issue 3 https://doi.org/10.1038/mt.2016.10	journal	March 2016
Using nanoBRET and CRISPR/Cas9 to monitor proximity to a genome-edited protein in real-time White, Carl W.; Vanyai, Hannah K.; See, Heng B. Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/s41598-017-03486-2	journal	June 2017
CRISPR/Cas9 editing reveals novel mechanisms of clustered microRNA regulation and function Lataniotis, Lazaros; Albrecht, Andreas; Kok, Fatma O. Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/s41598-017-09268-0	journal	August 2017
Simultaneous electroporation and dielectrophoresis in non-electrolytic micro/nano-electroporation Lyu, Chenang; Wang, Jianping; Powell-Palm, Matthew Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-018-20535-6	journal	February 2018
Efficient homology-directed gene editing by CRISPR/Cas9 in human stem and primary cells using tube electroporation Xu, Xiaoyun; Gao, Dongbing; Wang, Ping Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-018-30227-w	journal	August 2018
A comparison of inverted and upright laser-activated titanium nitride micropyramids for intracellular delivery Raun, Alexander; Saklayen, Nabiha; Zgrabik, Christine Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-018-33885-y	journal	October 2018
Co-expression of Cas9 and single-guided RNAs in Escherichia coli streamlines production of Cas9 ribonucleoproteins Qiao, Jie; Li, Wenqiang; Lin, Siyu Communications Biology, Vol. 2, Issue 1 https://doi.org/10.1038/s42003-019-0402-x	journal	May 2019
Understanding the diversity of genetic outcomes from CRISPR-Cas generated homology-directed repair Sansbury, Brett M.; Hewes, Amanda M.; Kmiec, Eric B. Communications Biology, Vol. 2, Issue 1 https://doi.org/10.1038/s42003-019-0705-y	journal	December 2019
Analyses of point mutation repair and allelic heterogeneity generated by CRISPR/Cas9 and single-stranded DNA oligonucleotides Bialk, Pawel; Sansbury, Brett; Rivera-Torres, Natalia Scientific Reports, Vol. 6, Issue 1 https://doi.org/10.1038/srep32681	journal	September 2016
A vector platform for the rapid and efficient engineering of stable complex transgenes Jäckel, Carsten; Nogueira, Melanie Schmitt; Ehni, Nadja Scientific Reports, Vol. 6, Issue 1 https://doi.org/10.1038/srep34365	journal	October 2016
Reprogramming MHC specificity by CRISPR-Cas9-assisted cassette exchange Kelton, William; Waindok, Ann Cathrin; Pesch, Theresa Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/srep45775	journal	April 2017
CRISPRs for Optimal Targeting: Delivery of CRISPR Components as DNA, RNA, and Protein into Cultured Cells and Single-Cell Embryos Kouranova, Evguenia; Forbes, Kevin; Zhao, Guojun Human Gene Therapy, Vol. 27, Issue 6 https://doi.org/10.1089/hum.2016.009	journal	June 2016
Discovery of an autoimmunity-associated IL2RA enhancer by unbiased targeting of transcriptional activation Simeonov, Dimitre R.; Gowen, Benjamin G.; Boontanrart, Mandy BioRxiv https://doi.org/10.1101/091843	posted_content	December 2016
Rewired signaling network in T cells expressing the chimeric antigen receptor (CAR) Dong, Rui; Libby, Kendra A.; Blaeschke, Franziska The EMBO Journal https://doi.org/10.1101/2019.12.19.883496	journal	December 2019
Mapping gene regulatory networks of primary CD4⁺ T cells using single-cell genomics and genome engineering Gate, Rachel E.; Kim, Min Cheol; Lu, Andrew BioRxiv https://doi.org/10.1101/678060	posted_content	June 2019
A CRISPR-Based Toolbox for Studying T Cell Signal Transduction Chi, Shen; Weiss, Arthur; Wang, Haopeng BioMed Research International, Vol. 2016 https://doi.org/10.1155/2016/5052369	journal	January 2016
Translational research—the need of a new bioethics approach Hostiuc, Sorin; Moldoveanu, Alin; Dascălu, Maria-Iuliana Journal of Translational Medicine, Vol. 14, Issue 1 https://doi.org/10.1186/s12967-016-0773-4	journal	January 2016
Strategies for precision modulation of gene expression by epigenome editing: an overview Laufer, Benjamin I.; Singh, Shiva M. Epigenetics & Chromatin, Vol. 8, Issue 1 https://doi.org/10.1186/s13072-015-0023-7	journal	September 2015
Targeting the latent reservoir to achieve functional HIV cure Cary, Daniele C.; Peterlin, B. Matija F1000Research, Vol. 5 https://doi.org/10.12688/f1000research.8109.1	journal	May 2016
A versatile system for rapid multiplex genome-edited CAR T cell generation Ren, Jiangtao; Zhang, Xuhua; Liu, Xiaojun Oncotarget, Vol. 8, Issue 10 https://doi.org/10.18632/oncotarget.15218	journal	February 2017
CRISPR-Cas9: A Revolutionary Tool for Cancer Modelling Torres-Ruiz, Raul; Rodriguez-Perales, Sandra International Journal of Molecular Sciences, Vol. 16, Issue 9 https://doi.org/10.3390/ijms160922151	journal	September 2015
RNA Interference in the Age of CRISPR: Will CRISPR Interfere with RNAi? Unniyampurath, Unnikrishnan; Pilankatta, Rajendra; Krishnan, Manoj International Journal of Molecular Sciences, Vol. 17, Issue 3 https://doi.org/10.3390/ijms17030291	journal	February 2016
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms Farboud, Behnom; Jarvis, Erin; Roth, Theodore L. Journal of Visualized Experiments, Issue 135 https://doi.org/10.3791/57350-v	journal	May 2018
CRISPR therapeutic tools for complex genetic disorders and cancer (Review) Baliou, Stella; Adamaki, Maria; Kyriakopoulos, Anthony International Journal of Oncology https://doi.org/10.3892/ijo.2018.4434	journal	June 2018
An Overview Of The Crispr-Based Genomic- And Epigenome-Editing System: Function, Applications, And Challenges Mojarrad, Majid; Bozorg Qomi, Saeed; Asghari, Amir Advanced Biomedical Research, Vol. 8, Issue 1 https://doi.org/10.4103/abr.abr_41_19	journal	January 2019
Efficient And Versatile Crispr Engineering Of Human Neurons In Culture To Model Neurological Disorders Shah, Ruth Rama; Cholewa-Waclaw, Justyna; Davies, Faith C. J. Zenodo https://doi.org/10.5281/zenodo.163342	text	January 2016
Correction: Multiplexed genetic engineering of human hematopoietic stem and progenitor cells using CRISPR/Cas9 and AAV6 Bak, Rasmus O.; Dever, Daniel P.; Reinisch, Andreas eLife, Vol. 7 https://doi.org/10.7554/elife.43690	journal	November 2018

Similar Records

Targeted delivery of CRISPR-Cas9 and transgenes enables complex immune cell engineering

Journal Article · Tue Jun 01 00:00:00 EDT 2021 · Cell Reports · OSTI ID:1221821

Hamilton, Jennifer R.; Tsuchida, Connor A.; Nguyen, David N.; +7 more

Establishment of a genome editing tool using CRISPR-Cas9 ribonucleoprotein complexes in the non-model plant pathogen Sphaerulina musiva

Journal Article · Fri Jul 21 00:00:00 EDT 2023 · Frontiers in Genome Editing · OSTI ID:1221821

Tannous, Joanna; Sawyer, Cole; Hassan, Md Mahmudul; +2 more

Targeted gene knock-in by homology-directed genome editing using Cas9 ribonucleoprotein and AAV donor delivery

Journal Article · Thu Mar 02 00:00:00 EST 2017 · Nucleic Acids Research · OSTI ID:1221821

Gaj, Thomas; Staahl, Brett T.; Rodrigues, Gonçalo M. C.; +4 more

Related Subjects

60 APPLIED LIFE SCIENCES
CRISPR/Cas9
genome engineering
Cas9 ribonucleoprotein
RNP primary
human T cells

Title: Generation of knock-in primary human T cells using Cas9 ribonucleoproteins

Citation Formats

References (24)

Cited By (166)

Similar Records

Related Subjects