Rational design of a split-Cas9 enzyme complex

Wright, Addison V.; Sternberg, Samuel H.; Taylor, David W.; Staahl, Brett T.; Bardales, Jorge A.; Kornfeld, Jack E.; Doudna, Jennifer A.

doi:10.1073/pnas.1501698112

Title: Rational design of a split-Cas9 enzyme complex

Journal Article · Mon Feb 23 00:00:00 EST 2015 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1501698112· OSTI ID:1221831

Wright, Addison V. ^[1]; Sternberg, Samuel H. ^[1]; Taylor, David W. ^[1]; Staahl, Brett T. ^[1]; Bardales, Jorge A. ^[1]; Kornfeld, Jack E. ^[1]; Doudna, Jennifer A. ^[2]

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

Cas9, an RNA-guided DNA endonuclease found in clustered regularly interspaced short palindromic repeats (CRISPR) bacterial immune systems, is a versatile tool for genome editing, transcriptional regulation, and cellular imaging applications. Structures of Streptococcus pyogenes Cas9 alone or bound to single-guide RNA (sgRNA) and target DNA revealed a bilobed protein architecture that undergoes major conformational changes upon guide RNA and DNA binding. To investigate the molecular determinants and relevance of the interlobe rearrangement for target recognition and cleavage, we designed a split-Cas9 enzyme in which the nuclease lobe and α-helical lobe are expressed as separate polypeptides. The lobes do not interact on their own, the sgRNA recruits them into a ternary complex that recapitulates the activity of full-length Cas9 and catalyzes site-specific DNA cleavage. The use of a modified sgRNA abrogates split-Cas9 activity by preventing dimerization, allowing for the development of an inducible dimerization system. We propose that split-Cas9 can act as a highly regulatable platform for genome-engineering applications.

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; (HHMI)

OSTI ID:: 1221831

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 112, Issue 10; 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: 205 works

Citation information provided by
Web of Science

References (30)

CRISPR-Cas Systems: Prokaryotes Upgrade to Adaptive Immunity Barrangou, Rodolphe; Marraffini, Luciano A. Molecular Cell, Vol. 54, Issue 2 https://doi.org/10.1016/j.molcel.2014.03.011	journal	April 2014
Unravelling the structural and mechanistic basis of CRISPR–Cas systems van der Oost, John; Westra, Edze R.; Jackson, Ryan N. Nature Reviews Microbiology, Vol. 12, Issue 7 https://doi.org/10.1038/nrmicro3279	journal	June 2014
The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli Sapranauskas, Rimantas; Gasiunas, Giedrius; Fremaux, Christophe Nucleic Acids Research, Vol. 39, Issue 21 https://doi.org/10.1093/nar/gkr606	journal	August 2011
A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity Jinek, M.; Chylinski, K.; Fonfara, I. Science, Vol. 337, Issue 6096, p. 816-821 https://doi.org/10.1126/science.1225829	journal	June 2012
Cas9 as a versatile tool for engineering biology Mali, Prashant; Esvelt, Kevin M.; Church, George M. Nature Methods, Vol. 10, Issue 10 https://doi.org/10.1038/nmeth.2649	journal	September 2013
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
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
Structures of Cas9 Endonucleases Reveal RNA-Mediated Conformational Activation Jinek, M.; Jiang, F.; Taylor, D. W. Science, Vol. 343, Issue 6176 https://doi.org/10.1126/science.1247997	journal	February 2014
Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA Nishimasu, Hiroshi; Ran, F. Ann; Hsu, Patrick D. Cell, Vol. 156, Issue 5 https://doi.org/10.1016/j.cell.2014.02.001	journal	February 2014
Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease Anders, Carolin; Niewoehner, Ole; Duerst, Alessia Nature, Vol. 513, Issue 7519 https://doi.org/10.1038/nature13579	journal	July 2014
Guide RNA Functional Modules Direct Cas9 Activity and Orthogonality Briner, Alexandra E.; Donohoue, Paul D.; Gomaa, Ahmed A. Molecular Cell, Vol. 56, Issue 2 https://doi.org/10.1016/j.molcel.2014.09.019	journal	October 2014
DNA targeting specificity of RNA-guided Cas9 nucleases Hsu, Patrick D.; Scott, David A.; Weinstein, Joshua A. Nature Biotechnology, Vol. 31, Issue 9, p. 827-832 https://doi.org/10.1038/nbt.2647	journal	July 2013
Split-protein systems: beyond binary protein–protein interactions Shekhawat, Sujan S.; Ghosh, Indraneel Current Opinion in Chemical Biology, Vol. 15, Issue 6 https://doi.org/10.1016/j.cbpa.2011.10.014	journal	December 2011
Optical control of mammalian endogenous transcription and epigenetic states Konermann, Silvana; Brigham, Mark D.; Trevino, Alexandro E. Nature, Vol. 500, Issue 7463 https://doi.org/10.1038/nature12466	journal	August 2013
Two- and three-input TALE-based AND logic computation in embryonic stem cells Lienert, Florian; Torella, Joseph P.; Chen, Jan-Hung Nucleic Acids Research, Vol. 41, Issue 21 https://doi.org/10.1093/nar/gkt758	journal	August 2013
Engineering the ABA Plant Stress Pathway for Regulation of Induced Proximity Liang, F. -S.; Ho, W. Q.; Crabtree, G. R. Science Signaling, Vol. 4, Issue 164 https://doi.org/10.1126/scisignal.2001449	journal	March 2011
Rapid blue-light–mediated induction of protein interactions in living cells Kennedy, Matthew J.; Hughes, Robert M.; Peteya, Leslie A. Nature Methods, Vol. 7, Issue 12 https://doi.org/10.1038/nmeth.1524	journal	October 2010
Structures of the RNA-guided surveillance complex from a bacterial immune system Wiedenheft, Blake; Lander, Gabriel C.; Zhou, Kaihong Nature, Vol. 477, Issue 7365 https://doi.org/10.1038/nature10402	journal	September 2011
Crystal structure of the CRISPR RNA-guided surveillance complex from Escherichia coli Jackson, R. N.; Golden, S. M.; van Erp, P. B. G. Science, Vol. 345, Issue 6203 https://doi.org/10.1126/science.1256328	journal	August 2014
Crystal structure of a CRISPR RNA-guided surveillance complex bound to a ssDNA target Mulepati, S.; Heroux, A.; Bailey, S. Science, Vol. 345, Issue 6203 https://doi.org/10.1126/science.1256996	journal	August 2014
Automated molecular microscopy: The new Leginon system Suloway, Christian; Pulokas, James; Fellmann, Denis Journal of Structural Biology, Vol. 151, Issue 1 https://doi.org/10.1016/j.jsb.2005.03.010	journal	July 2005
Complete subunit architecture of the proteasome regulatory particle Lander, Gabriel C.; Estrin, Eric; Matyskiela, Mary E. Nature, Vol. 482, Issue 7384 https://doi.org/10.1038/nature10774	journal	January 2012
A New Generation of the IMAGIC Image Processing System van Heel, Marin; Harauz, George; Orlova, Elena V. Journal of Structural Biology, Vol. 116, Issue 1 https://doi.org/10.1006/jsbi.1996.0004	journal	January 1996
Targeted genome editing in human cells with zinc finger nucleases constructed via modular assembly Kim, H. J.; Lee, H. J.; Kim, H. Genome Research, Vol. 19, Issue 7 https://doi.org/10.1101/gr.089417.108	journal	May 2009
Double Nicking by RNA-Guided CRISPR Cas9 for Enhanced Genome Editing Specificity Ran, F. Ann; Hsu, Patrick D.; Lin, Chie-Yu Cell, Vol. 154, Issue 6, p. 1380-1389 https://doi.org/10.1016/j.cell.2013.08.021	journal	September 2013
Double Nicking by RNA-Guided CRISPR Cas9 for Enhanced Genome Editing Specificity Ran, F. Ann; Hsu, Patrick D.; Lin, Chie-Yu Cell, Vol. 155, Issue 2 https://doi.org/10.1016/j.cell.2013.09.040	journal	October 2013
Optical control of mammalian endogenous transcription and epigenetic states Konermann, Silvana; Brigham, Mark D.; Trevino, Alexandro Nature https://doi.org/10.1038/nature20002	journal	June 2016
Structures of Cas9 endonucleases reveal RNA-mediated conformational activation Jinek, Martin; Jiang, Fuguo; Taylor, David W. American Association for the Advancement of Science https://doi.org/10.5167/uzh-96990	text	January 2014
Optical control of mammalian endogenous transcription and epigenetic states Konermann, Silvana; Brigham, Mark D.; Trevino, Alexandro Nature https://doi.org/10.1038/nature20003	journal	June 2016
Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease Carolin, Anders,; Ole, Niewoehner,; Alessia, Duerst, Nature Publishing Group https://doi.org/10.5167/uzh-99826	text	January 2014

Cited By (79)

Distinct timescales of RNA regulators enable the construction of a genetic pulse generator Westbrook, Alexandra; Tang, Xun; Marshall, Ryan Biotechnology and Bioengineering, Vol. 116, Issue 5 https://doi.org/10.1002/bit.26918	journal	February 2019
CRISPR-Cas9 in genome editing: Its function and medical applications: KHADEMPAR et al. Khadempar, Saedeh; Familghadakchi, Shokoufeh; Motlagh, Roozbeh Akbari Journal of Cellular Physiology, Vol. 234, Issue 5 https://doi.org/10.1002/jcp.27476	journal	October 2018
Engineering large animal models of human disease Whitelaw, C. Bruce A.; Sheets, Timothy P.; Lillico, Simon G. The Journal of Pathology, Vol. 238, Issue 2 https://doi.org/10.1002/path.4648	journal	November 2015
Heat‐shock‐inducible CRISPR/Cas9 system generates heritable mutations in rice Nandy, Soumen; Pathak, Bhuvan; Zhao, Shan Plant Direct, Vol. 3, Issue 5 https://doi.org/10.1002/pld3.145	journal	May 2019
Cas9 versus Cas12a/Cpf1: Structure-function comparisons and implications for genome editing Swarts, Daan C.; Jinek, Martin Wiley Interdisciplinary Reviews: RNA, Vol. 9, Issue 5 https://doi.org/10.1002/wrna.1481	journal	May 2018
Using CRISPR-Cas9 Genome Editing to Enhance Cell Based Therapies for the Treatment of Diabetes Mellitus Krentz, Nicole A. J.; Lynn, Francis C. Genome Editing https://doi.org/10.1007/978-3-319-34148-4_8	book	August 2016
The expanding footprint of CRISPR/Cas9 in the plant sciences Schaeffer, Scott M.; Nakata, Paul A. Plant Cell Reports, Vol. 35, Issue 7 https://doi.org/10.1007/s00299-016-1987-x	journal	April 2016
CRISPR-Cas orthologues and variants: optimizing the repertoire, specificity and delivery of genome engineering tools Cebrian-Serrano, Alberto; Davies, Benjamin Mammalian Genome, Vol. 28, Issue 7-8 https://doi.org/10.1007/s00335-017-9697-4	journal	June 2017
Gene targeting, genome editing: from Dolly to editors Tan, Wenfang; Proudfoot, Chris; Lillico, Simon G. Transgenic Research, Vol. 25, Issue 3 https://doi.org/10.1007/s11248-016-9932-x	journal	February 2016
Development of CRISPR-Cas systems for genome editing and beyond Zhang, F. Quarterly Reviews of Biophysics, Vol. 52 https://doi.org/10.1017/s0033583519000052	journal	January 2019
Conformational control of DNA target cleavage by CRISPR–Cas9 Sternberg, Samuel H.; LaFrance, Benjamin; Kaplan, Matias Nature, Vol. 527, Issue 7576 https://doi.org/10.1038/nature15544	journal	October 2015
Structural basis of CRISPR–SpyCas9 inhibition by an anti-CRISPR protein Dong, De; Guo, Minghui; Wang, Sihan Nature, Vol. 546, Issue 7658 https://doi.org/10.1038/nature22377	journal	April 2017
Enhanced proofreading governs CRISPR–Cas9 targeting accuracy Chen, Janice S.; Dagdas, Yavuz S.; Kleinstiver, Benjamin P. Nature, Vol. 550, Issue 7676 https://doi.org/10.1038/nature24268	journal	September 2017
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
Rapid and tunable method to temporally control gene editing based on conditional Cas9 stabilization Senturk, Serif; Shirole, Nitin H.; Nowak, Dawid G. Nature Communications, Vol. 8, Issue 1 https://doi.org/10.1038/ncomms14370	journal	February 2017
In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni Kim, Eunji; Koo, Taeyoung; Park, Sung Wook Nature Communications, Vol. 8, Issue 1 https://doi.org/10.1038/ncomms14500	journal	February 2017
Hit and go CAS9 delivered through a lentiviral based self-limiting circuit Petris, Gianluca; Casini, Antonio; Montagna, Claudia Nature Communications, Vol. 8, Issue 1 https://doi.org/10.1038/ncomms15334	journal	May 2017
Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display Shechner, David M.; Hacisuleyman, Ezgi; Younger, Scott T. Nature Methods, Vol. 12, Issue 7 https://doi.org/10.1038/nmeth.3433	journal	June 2015
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
A multifunctional AAV–CRISPR–Cas9 and its host response Chew, Wei Leong; Tabebordbar, Mohammadsharif; Cheng, Jason K. W. Nature Methods, Vol. 13, Issue 10 https://doi.org/10.1038/nmeth.3993	journal	September 2016
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
Defining and improving the genome-wide specificities of CRISPR–Cas9 nucleases Tsai, Shengdar Q.; Joung, J. Keith Nature Reviews Genetics, Vol. 17, Issue 5 https://doi.org/10.1038/nrg.2016.28	journal	April 2016
A thermostable Cas9 with increased lifetime in human plasma Harrington, Lucas B.; Paez-Espino, David; Staahl, Brett T. Nature Communications, Vol. 8, Issue 1 https://doi.org/10.1038/s41467-017-01408-4	journal	November 2017
Genome-wide profiling of adenine base editor specificity by EndoV-seq Liang, Puping; Xie, Xiaowei; Zhi, Shengyao Nature Communications, Vol. 10, Issue 1 https://doi.org/10.1038/s41467-018-07988-z	journal	January 2019
CRISPR-Switch regulates sgRNA activity by Cre recombination for sequential editing of two loci Chylinski, Krzysztof; Hubmann, Maria; Hanna, Ruth E. Nature Communications, Vol. 10, Issue 1 https://doi.org/10.1038/s41467-019-13403-y	journal	November 2019
Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated viruses Levy, Jonathan M.; Yeh, Wei-Hsi; Pendse, Nachiket Nature Biomedical Engineering, Vol. 4, Issue 1 https://doi.org/10.1038/s41551-019-0501-5	journal	January 2020
CRISPR–Cas9 genome editing in human cells occurs via the Fanconi anemia pathway Richardson, Chris D.; Kazane, Katelynn R.; Feng, Sharon J. Nature Genetics, Vol. 50, Issue 8 https://doi.org/10.1038/s41588-018-0174-0	journal	July 2018
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
A split CRISPR–Cpf1 platform for inducible genome editing and gene activation Nihongaki, Yuta; Otabe, Takahiro; Ueda, Yoshibumi Nature Chemical Biology, Vol. 15, Issue 9 https://doi.org/10.1038/s41589-019-0338-y	journal	August 2019
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
CRISPR RNA-guided autonomous delivery of Cas9 Wilkinson, Royce A.; Martin, Coleman; Nemudryi, Artem A. Nature Structural & Molecular Biology, Vol. 26, Issue 1 https://doi.org/10.1038/s41594-018-0173-y	journal	December 2018
Trans-spliced Cas9 allows cleavage of HBB and CCR5 genes in human cells using compact expression cassettes Fine, Eli J.; Appleton, Caleb M.; White, Douglas E. Scientific Reports, Vol. 5, Issue 1 https://doi.org/10.1038/srep10777	journal	July 2015
Addressing biological uncertainties in engineering gene circuits Zhang, Carolyn; Tsoi, Ryan; You, Lingchong Integrative Biology, Vol. 8, Issue 4 https://doi.org/10.1039/c5ib00275c	journal	January 2016
CRISPR-Cas9 conformational activation as elucidated from enhanced molecular simulations Palermo, Giulia; Miao, Yinglong; Walker, Ross C. Proceedings of the National Academy of Sciences, Vol. 114, Issue 28 https://doi.org/10.1073/pnas.1707645114	journal	June 2017
Genome editing in crop improvement: Present scenario and future prospects Miglani, Gurbachan S. Journal of Crop Improvement, Vol. 31, Issue 4 https://doi.org/10.1080/15427528.2017.1333192	journal	June 2017
Plant gene expression control using genome- and epigenome-editing technologies Miglani, Gurbachan S.; Kaur, Anroop; Kaur, Lovepreet Journal of Crop Improvement, Vol. 34, Issue 1 https://doi.org/10.1080/15427528.2019.1678541	journal	October 2019
Recent advances in developing and applying biosensors for synthetic biology Jones, Krysten A.; Zinkus-Boltz, Julia; Dickinson, Bryan C. Nano Futures, Vol. 3, Issue 4 https://doi.org/10.1088/2399-1984/ab4b78	journal	October 2019
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
Development of an intein-mediated split–Cas9 system for gene therapy Truong, Dong-Jiunn Jeffery; Kühner, Karin; Kühn, Ralf Nucleic Acids Research, Vol. 43, Issue 13 https://doi.org/10.1093/nar/gkv601	journal	June 2015
Kinetics of the CRISPR-Cas9 effector complex assembly and the role of 3′-terminal segment of guide RNA Mekler, Vladimir; Minakhin, Leonid; Semenova, Ekaterina Nucleic Acids Research, Vol. 44, Issue 6 https://doi.org/10.1093/nar/gkw138	journal	March 2016
An easy and efficient inducible CRISPR/Cas9 platform with improved specificity for multiple gene targeting Cao, Jian; Wu, Lizhen; Zhang, Shang-Min Nucleic Acids Research https://doi.org/10.1093/nar/gkw660	journal	July 2016
Engineered dCas9 with reduced toxicity in bacteria: implications for genetic circuit design Zhang, Shuyi; Voigt, Christopher A. Nucleic Acids Research https://doi.org/10.1093/nar/gky884	journal	October 2018
Cell-specific CRISPR–Cas9 activation by microRNA-dependent expression of anti-CRISPR proteins Hoffmann, Mareike D.; Aschenbrenner, Sabine; Grosse, Stefanie Nucleic Acids Research, Vol. 47, Issue 13 https://doi.org/10.1093/nar/gkz271	journal	April 2019
All-in-One Adeno-associated Virus Delivery and Genome Editing by Neisseria meningitidis Cas9 in vivo Ibraheim, Raed; Song, Chun-Qing; Mir, Aamir bioRxiv https://doi.org/10.1101/295055	posted_content	May 2018
Differential efficacies of Cas nucleases on microsatellites involved in human disorders and associated off-target mutations Poggi, Lucie; Emmenegger, Lisa; Descorps-Declère, Stéphane bioRxiv https://doi.org/10.1101/857714	posted_content	November 2019
A Model for Resource Competition in CRISPR-Mediated Gene Repression Chen, Pin-Yi; Qian, Yili; Del Vecchio, Domitilla 2018 IEEE Conference on Decision and Control (CDC) https://doi.org/10.1109/cdc.2018.8619016	conference	December 2018
New variants of CRISPR RNA-guided genome editing enzymes Murovec, Jana; Pirc, Žan; Yang, Bing Plant Biotechnology Journal, Vol. 15, Issue 8 https://doi.org/10.1111/pbi.12736	journal	May 2017
Unexpected evolutionary benefit to phages imparted by bacterial CRISPR-Cas9 Tao, Pan; Wu, Xiaorong; Rao, Venigalla Science Advances, Vol. 4, Issue 2 https://doi.org/10.1126/sciadv.aar4134	journal	February 2018
CRISPR/CAS9, the king of genome editing tools Bannikov, A. V.; Lavrov, A. V. Molecular Biology, Vol. 51, Issue 4 https://doi.org/10.1134/s0026893317040033	journal	July 2017
Protein Engineering Strategies to Expand CRISPR-Cas9 Applications Ribeiro, Lucas F.; Ribeiro, Liliane F. C.; Barreto, Matheus Q. International Journal of Genomics, Vol. 2018 https://doi.org/10.1155/2018/1652567	journal	August 2018
Novel Epigenetic Techniques Provided by the CRISPR/Cas9 System Xie, Nina; Zhou, Yafang; Sun, Qiying Stem Cells International, Vol. 2018 https://doi.org/10.1155/2018/7834175	journal	July 2018
Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage Kulcsár, Péter István; Tálas, András; Huszár, Krisztina Genome Biology, Vol. 18, Issue 1 https://doi.org/10.1186/s13059-017-1318-8	journal	October 2017
All-in-one adeno-associated virus delivery and genome editing by Neisseria meningitidis Cas9 in vivo Ibraheim, Raed; Song, Chun-Qing; Mir, Aamir Genome Biology, Vol. 19, Issue 1 https://doi.org/10.1186/s13059-018-1515-0	journal	September 2018
From fiction to science: clinical potentials and regulatory considerations of gene editing Schacker, Maria; Seimetz, Diane Clinical and Translational Medicine, Vol. 8, Issue 1 https://doi.org/10.1186/s40169-019-0244-7	journal	October 2019
Staphylococcus aureus Cas9 is a multiple-turnover enzyme Yourik, Paul; Fuchs, Ryan T.; Mabuchi, Megumu RNA, Vol. 25, Issue 1 https://doi.org/10.1261/rna.067355.118	journal	October 2018
Genetic and epigenetic control of gene expression by CRISPR–Cas systems Lo, Albert; Qi, Lei F1000Research, Vol. 6 https://doi.org/10.12688/f1000research.11113.1	journal	January 2017
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
A new age in functional genomics using CRISPR/Cas9 in arrayed library screening Agrotis, Alexander; Ketteler, Robin Frontiers in Genetics, Vol. 6 https://doi.org/10.3389/fgene.2015.00300	journal	September 2015
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
CRISPR/Cas9 for Cancer Therapy: Hopes and Challenges Martinez-Lage, Marta; Puig-Serra, Pilar; Menendez, Pablo Biomedicines, Vol. 6, Issue 4 https://doi.org/10.3390/biomedicines6040105	journal	November 2018
CRISPR-Cas9 conformational activation as elucidated from enhanced molecular simulations Giulia, Palermo,; Yinglong, Miao,; C., Walker, Ross National Academy of Sciences https://doi.org/10.5167/uzh-138566	text	January 2017
Crispr/Cas9 - КОРОЛЬ ГЕНОМНОГО РЕДАКТИРОВАНИЯ, "Молекулярная биология" Банников, А. В.; Лавров, А. В. Молекулярная биология, Issue 4 https://doi.org/10.7868/S0026898417040036	journal	January 2017
Cas9 versus Cas12a/Cpf1: Structure-function comparisons and implications for genome editing Swarts, Daan C.; Jinek, Martin Wiley-Blackwell Publishing, Inc. https://doi.org/10.5167/uzh-168880	text	January 2018
Differential efficacies of Cas nucleases on microsatellites involved in human disorders and associated off-target mutations Poggi, Lucie; Emmenegger, Lisa; Descorps-Declère, Stéphane Nucleic Acids Research, Vol. 49, Issue 14 https://doi.org/10.1093/nar/gkab569	journal	July 2021
Allosteric inhibition of CRISPR-Cas9 by bacteriophage-derived peptides Cui, Yan-ru; Wang, Shao-jie; Chen, Jun Genome Biology, Vol. 21, Issue 1 https://doi.org/10.1186/s13059-020-01956-x	journal	February 2020
Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage István Kulcsér, Péter; Tálas, András; Huszár, Krisztina Genome Biology https://doi.org/10.1101/187898	posted_content	September 2017
Programmable Genome Editing Tools and their Regulation for Efficient Genome Engineering Guha, Tuhin Kumar; Wai, Alvan; Hausner, Georg Computational and Structural Biotechnology Journal, Vol. 15 https://doi.org/10.1016/j.csbj.2016.12.006	journal	January 2017
HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control Zhao, Chen; Zhao, Yingze; Zhang, Jingfang Molecular Therapy - Nucleic Acids, Vol. 13 https://doi.org/10.1016/j.omtn.2018.08.022	journal	December 2018
New and emerging uses of CRISPR/Cas9 to genetically manipulate apicomplexan parasites Di Cristina, Manlio; Carruthers, Vern B. Parasitology, Vol. 145, Issue 9 https://doi.org/10.1017/s003118201800001x	journal	February 2018
Striking Plasticity of CRISPR-Cas9 and Key Role of Non-target DNA, as Revealed by Molecular Simulations Palermo, Giulia; Miao, Yinglong; Walker, Ross C. ACS Central Science, Vol. 2, Issue 10 https://doi.org/10.1021/acscentsci.6b00218	journal	September 2016
Ligand-binding domains of nuclear receptors facilitate tight control of split CRISPR activity Nguyen, Duy P.; Miyaoka, Yuichiro; Gilbert, Luke A. Nature Communications, Vol. 7, Issue 1 https://doi.org/10.1038/ncomms12009	journal	July 2016
Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells Ma, Dacheng; Peng, Shuguang; Xie, Zhen Nature Communications, Vol. 7, Issue 1 https://doi.org/10.1038/ncomms13056	journal	October 2016
Improved Cas9 activity by specific modifications of the tracrRNA Scott, Tristan; Urak, Ryan; Soemardy, Citradewi Scientific Reports, Vol. 9, Issue 1 https://doi.org/10.1038/s41598-019-52616-5	journal	November 2019
In vivoblunt-end cloning through CRISPR/Cas9-facilitated non-homologous end-joining Geisinger, Jonathan M.; Turan, Sören; Hernandez, Sophia Nucleic Acids Research, Vol. 44, Issue 8 https://doi.org/10.1093/nar/gkv1542	journal	January 2016
Precision cancer mouse models through genome editing with CRISPR-Cas9 Mou, Haiwei; Kennedy, Zachary; Anderson, Daniel G. Genome Medicine, Vol. 7, Issue 1 https://doi.org/10.1186/s13073-015-0178-7	journal	June 2015
Brain tumor modeling using the CRISPR/Cas9 system: state of the art and view to the future Mao, Xiao-Yuan; Dai, Jin-Xiang; Zhou, Hong-Hao Oncotarget, Vol. 7, Issue 22 https://doi.org/10.18632/oncotarget.8075	journal	March 2016
Genome Engineering with TALE and CRISPR Systems in Neuroscience Lee, Han B.; Sundberg, Brynn N.; Sigafoos, Ashley N. Frontiers in Genetics, Vol. 7 https://doi.org/10.3389/fgene.2016.00047	journal	April 2016
Genome editing: A perspective on the application of CRISPR/Cas9 to study human diseases (Review) Rodr�guez‑Rodr�guez, Diana; Ram�rez‑Sol�s, Ramiro; Garza‑Elizondo, Mario International Journal of Molecular Medicine https://doi.org/10.3892/ijmm.2019.4112	journal	February 2019
The length of guide RNA and target DNA heteroduplex effects on CRISPR/Cas9 mediated genome editing efficiency in porcine cells Lv, Jiawei; Wu, Shuang; Wei, Renyue Journal of Veterinary Science, Vol. 20, Issue 3 https://doi.org/10.4142/jvs.2019.20.e23	journal	January 2019

Similar Records

Development of a CRISPR/Cas9 System for Methylococcus capsulatus In Vivo Gene Editing

Journal Article · Fri Mar 29 00:00:00 EDT 2019 · Applied and Environmental Microbiology · OSTI ID:1221831

Tapscott, Timothy; Guarnieri, Michael T.; Henard, Calvin A.; +1 more

Use of anti-CRISPR protein AcrIIA4 as a capture ligand for CRISPR/Cas9 detection

Journal Article · Sun Sep 15 00:00:00 EDT 2019 · Biosensors and Bioelectronics · OSTI ID:1221831

Johnston, Robert K.; Seamon, Kyle J.; Saada, Edwin A.; +4 more

CRISPR/Cas9-Mediated Phage Resistance Is Not Impeded by the DNA Modifications of Phage T4

Journal Article · Mon Jun 02 00:00:00 EDT 2014 · PLoS ONE · OSTI ID:1221831

Yaung, Stephanie J.; Esvelt, Kevin M.; Church, George M.

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CRISPR-Cas9
genome engineering
split enzyme

Title: Rational design of a split-Cas9 enzyme complex

Citation Formats

References (30)

Cited By (79)

Similar Records

Related Subjects