Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice

Jiang, Wenzhi; Zhou, Huanbin; Bi, Honghao; Fromm, Michael; Yang, Bing; Weeks, Donald P.

doi:10.1093/nar/gkt780

Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice

Journal Article · Sat Aug 31 00:00:00 EDT 2013 · Nucleic Acids Research

DOI:https://doi.org/10.1093/nar/gkt780· OSTI ID:1904805

Jiang, Wenzhi ^[1]; Zhou, Huanbin ^[2]; Bi, Honghao ^[2]; Fromm, Michael ^[3]; Yang, Bing ^[2]; Weeks, Donald P. ^[3]

Univ. of Nebraska, Lincoln, NE (United States); Univ. of Nebraska, Lincoln, NE (United States)
Iowa State Univ., Ames, IA (United States)
Univ. of Nebraska, Lincoln, NE (United States)

The type II CRISPR/Cas system from Streptococcus pyogenes and its simplified derivative, the Cas9/single guide RNA (sgRNA) system, have emerged as potent new tools for targeted gene knockout in bacteria, yeast, fruit fly, zebrafish and human cells. Here, we describe adaptations of these systems leading to successful expression of the Cas9/sgRNA system in two dicot plant species, Arabidopsis and tobacco, and two monocot crop species, rice and sorghum. Agrobacterium tumefaciens was used for delivery of genes encoding Cas9, sgRNA and a non-functional, mutant green fluorescence protein (GFP) to Arabidopsis and tobacco. The mutant GFP gene contained target sites in its 5' coding regions that were successfully cleaved by a CAS9/sgRNA complex that, along with error-prone DNA repair, resulted in creation of functional GFP genes. DNA sequencing confirmed Cas9/sgRNA-mediated mutagenesis at the target site. Rice protoplast cells transformed with Cas9/sgRNA constructs targeting the promoter region of the bacterial blight susceptibility genes, OsSWEET14 and OsSWEET11, were confirmed by DNA sequencing to contain mutated DNA sequences at the target sites. Successful demonstration of the Cas9/sgRNA system in model plant and crop species bodes well for its near-term use as a facile and powerful means of plant genetic engineering for scientific and agricultural applications.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of California, San Diego, CA (United States); Univ. of Nebraska, Lincoln, NE (United States)

Sponsoring Organization:: National Science Foundation (NSF); USDOE Office of Energy Efficiency and Renewable Energy (EERE)

Grant/Contract Number:: EE0001052; EE0003373

OSTI ID:: 1904805

Journal Information:: Nucleic Acids Research, Journal Name: Nucleic Acids Research Journal Issue: 20 Vol. 41; ISSN 0305-1048

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United States

Language:: English

References (23)

Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants Christensen, Alan H.; Quail, Peter H. Transgenic Research, Vol. 5, Issue 3 https://doi.org/10.1007/bf01969712	journal	May 1996
Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components Porebski, Sue; Bailey, L. Grant; Baum, Bernard R. Plant Molecular Biology Reporter, Vol. 15, Issue 1 https://doi.org/10.1007/bf02772108	journal	March 1997
Rapid and reproducible Agrobacterium-mediated transformation of sorghum Howe, Arlene; Sato, Shirley; Dweikat, Ismail Plant Cell Reports, Vol. 25, Issue 8 https://doi.org/10.1007/s00299-005-0081-6	journal	March 2006
High frequency targeting of genes to specific sites in the mammalian genome Thomas, K. Cell, Vol. 44, Issue 3 https://doi.org/10.1016/0092-8674(86)90463-0	journal	February 1986
One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering Wang, Haoyi; Yang, Hui; Shivalila, Chikdu S. Cell, Vol. 153, Issue 4, p. 910-918 https://doi.org/10.1016/j.cell.2013.04.025	journal	May 2013
ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering Gaj, Thomas; Gersbach, Charles A.; Barbas, Carlos F. Trends in Biotechnology, Vol. 31, Issue 7, p. 397-405 https://doi.org/10.1016/j.tibtech.2013.04.004	journal	July 2013
Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos Chang, Nannan; Sun, Changhong; Gao, Lu Cell Research, Vol. 23, Issue 4 https://doi.org/10.1038/cr.2013.45	journal	March 2013
RNA-guided genetic silencing systems in bacteria and archaea Wiedenheft, Blake; Sternberg, Samuel H.; Doudna, Jennifer A. Nature, Vol. 482, Issue 7385, p. 331-338 https://doi.org/10.1038/nature10886	journal	February 2012
High-efficiency TALEN-based gene editing produces disease-resistant rice Li, Ting; Liu, Bo; Spalding, Martin H. Nature Biotechnology, Vol. 30, Issue 5 https://doi.org/10.1038/nbt.2199	journal	May 2012
A ligation-independent cloning technique for high-throughput assembly of transcription activator–like effector genes Schmid-Burgk, Jonathan L.; Schmidt, Tobias; Kaiser, Vera Nature Biotechnology, Vol. 31, Issue 1 https://doi.org/10.1038/nbt.2460	journal	December 2012
Targeted genome modification of crop plants using a CRISPR-Cas system Shan, Qiwei; Wang, Yanpeng; Li, Jun Nature Biotechnology, Vol. 31, Issue 8 https://doi.org/10.1038/nbt.2650	journal	August 2013
Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease Nekrasov, Vladimir; Staskawicz, Brian; Weigel, Detlef Nature Biotechnology, Vol. 31, Issue 8 https://doi.org/10.1038/nbt.2655	journal	August 2013
Engineering polydactyl zinc-finger transcription factors Beerli, Roger R.; Barbas, Carlos F. Nature Biotechnology, Vol. 20, Issue 2 https://doi.org/10.1038/nbt0202-135	journal	February 2002
High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases Chen, Fuqiang; Pruett-Miller, Shondra M.; Huang, Yuping Nature Methods, Vol. 8, Issue 9 https://doi.org/10.1038/nmeth.1653	journal	July 2011
RNA-guided gene editing de Souza, Natalie Nature Methods, Vol. 10, Issue 3 https://doi.org/10.1038/nmeth.2389	journal	February 2013
A revolution coming to a classic model organism Grunwald, David Jonah Nature Methods, Vol. 10, Issue 4 https://doi.org/10.1038/nmeth.2415	journal	March 2013
CRISPR silencing de Souza, Natalie Nature Methods, Vol. 10, Issue 5 https://doi.org/10.1038/nmeth.2466	journal	April 2013
Targeted gene addition into a specified location in the human genome using designed zinc finger nucleases Moehle, E. A.; Rock, J. M.; Lee, Y. -L. Proceedings of the National Academy of Sciences, Vol. 104, Issue 9 https://doi.org/10.1073/pnas.0611478104	journal	February 2007
Efficient TALEN-mediated gene knockout in livestock Carlson, Daniel F.; Tan, Wenfang; Lillico, Simon G. Proceedings of the National Academy of Sciences, Vol. 109, Issue 43 https://doi.org/10.1073/pnas.1211446109	journal	October 2012
Modularly assembled designer TAL effector nucleases for targeted gene knockout and gene replacement in eukaryotes Li, Ting; Huang, Sheng; Zhao, Xuefeng Nucleic Acids Research, Vol. 39, Issue 14 https://doi.org/10.1093/nar/gkr188	journal	March 2011
A Simple Cipher Governs DNA Recognition by TAL Effectors Moscou, M. J.; Bogdanove, A. J. Science, Vol. 326, Issue 5959 https://doi.org/10.1126/science.1178817	journal	October 2009
DNA Double-Strand Break Repair: All's Well that Ends Well Wyman, Claire; Kanaar, Roland Annual Review of Genetics, Vol. 40, Issue 1 https://doi.org/10.1146/annurev.genet.40.110405.090451	journal	December 2006
A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes Zhang, Yang; Su, Jianbin; Duan, Shan Plant Methods, Vol. 7, Issue 1 https://doi.org/10.1186/1746-4811-7-30	journal	January 2011

Cited By (67)

Considerations in adapting CRISPR/Cas9 in nongenetic model plant systems Shan, Shengchen; Soltis, Pamela S.; Soltis, Douglas E. Applications in Plant Sciences, Vol. 8, Issue 1 https://doi.org/10.1002/aps3.11314	journal	January 2020
Sorghum stem aerenchyma formation is regulated by SbNAC_D during internode development Casto, Anna L.; McKinley, Brian A.; Yu, Ka Man Jasmine Plant Direct, Vol. 2, Issue 11 https://doi.org/10.1002/pld3.85	journal	November 2018
Comparison of CRISPR/Cas9 expression constructs for efficient targeted mutagenesis in rice Mikami, Masafumi; Toki, Seiichi; Endo, Masaki Plant Molecular Biology, Vol. 88, Issue 6 https://doi.org/10.1007/s11103-015-0342-x	journal	July 2015
Optimizing the DNA Donor Template for Homology-Directed Repair of Double-Strand Breaks Song, Fei; Stieger, Knut Molecular Therapy - Nucleic Acids, Vol. 7 https://doi.org/10.1016/j.omtn.2017.02.006	journal	June 2017
Safeguarding CRISPR-Cas9 gene drives in yeast DiCarlo, James E.; Chavez, Alejandro; Dietz, Sven L. Nature Biotechnology, Vol. 33, Issue 12 https://doi.org/10.1038/nbt.3412	journal	November 2015
Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus Lu, Shan; Shen, Xiaorui; Chen, Baoshan Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/s41598-017-03233-7	journal	June 2017
Validation of microinjection methods for generating knockout mice by CRISPR/Cas-mediated genome engineering Horii, Takuro; Arai, Yuji; Yamazaki, Miho Scientific Reports, Vol. 4, Issue 1 https://doi.org/10.1038/srep04513	journal	March 2014
Generation of inheritable and “transgene clean” targeted genome-modified rice in later generations using the CRISPR/Cas9 system Xu, Rong-Fang; Li, Hao; Qin, Rui-Ying Scientific Reports, Vol. 5, Issue 1 https://doi.org/10.1038/srep11491	journal	June 2015
CRMAGE: CRISPR Optimized MAGE Recombineering Ronda, Carlotta; Pedersen, Lasse Ebdrup; Sommer, Morten O. A. Scientific Reports, Vol. 6, Article No. 19452 https://doi.org/10.1038/srep19452	journal	January 2016
CRISPR/Cas9-mediated efficient and heritable targeted mutagenesis in tomato plants in the first and later generations Pan, Changtian; Ye, Lei; Qin, Li Scientific Reports, Vol. 6, Issue 1 https://doi.org/10.1038/srep24765	journal	April 2016
A high-efficiency CRISPR/Cas9 system for targeted mutagenesis in Cotton (Gossypium hirsutum L.) Li, Chao; Unver, Turgay; Zhang, Baohong Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/srep43902	journal	March 2017
Now for the hard ones: is there a limit on CRISPR genome editing in crops? Botella, Jose R. Journal of Experimental Botany, Vol. 70, Issue 3 https://doi.org/10.1093/jxb/erz007	journal	January 2019
Efficient chromosomal gene modification with CRISPR/cas9 and PCR-based homologous recombination donors in cultured Drosophila cells Bottcher, R.; Hollmann, M.; Merk, K. Nucleic Acids Research, Vol. 42, Issue 11 https://doi.org/10.1093/nar/gku289	journal	April 2014
CRISPR-Cas: an efficient tool for genome engineering of virulent bacteriophages Martel, Bruno; Moineau, Sylvain Nucleic Acids Research, Vol. 42, Issue 14 https://doi.org/10.1093/nar/gku628	journal	July 2014
An Agrobacterium -delivered CRISPR/Cas9 system for high-frequency targeted mutagenesis in maize Char, Si Nian; Neelakandan, Anjanasree K.; Nahampun, Hartinio Plant Biotechnology Journal, Vol. 15, Issue 2 https://doi.org/10.1111/pbi.12611	journal	September 2016
Significant enhancement of fatty acid composition in seeds of the allohexaploid, Camelina sativa , using CRISPR/Cas9 gene editing Jiang, Wen Zhi; Henry, Isabelle M.; Lynagh, Peter G. Plant Biotechnology Journal, Vol. 15, Issue 5 https://doi.org/10.1111/pbi.12663	journal	January 2017
Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration Lin, Choun-Sea; Hsu, Chen-Tran; Yang, Ling-Hung Plant Biotechnology Journal, Vol. 16, Issue 7 https://doi.org/10.1111/pbi.12870	journal	January 2018
Developing a flexible, high-efficiency Agrobacterium -mediated sorghum transformation system with broad application Che, Ping; Anand, Ajith; Wu, Emily Plant Biotechnology Journal, Vol. 16, Issue 7 https://doi.org/10.1111/pbi.12879	journal	February 2018
Efficient genome editing of wild strawberry genes, vector development and validation Zhou, Junhui; Wang, Guoming; Liu, Zhongchi Plant Biotechnology Journal, Vol. 16, Issue 11 https://doi.org/10.1111/pbi.12922	journal	April 2018
A new rice breeding method: CRISPR/Cas9 system editing of the Xa13 promoter to cultivate transgene‐free bacterial blight‐resistant rice Li, Changyan; Li, Wei; Zhou, Zaihui Plant Biotechnology Journal, Vol. 18, Issue 2 https://doi.org/10.1111/pbi.13217	journal	August 2019
Gene targeting using the Agrobacterium tumefaciens-mediated CRISPR-Cas system in rice Xu, Rongfang; Li, Hao; Qin, Ruiying Rice, Vol. 7, Issue 1 https://doi.org/10.1186/s12284-014-0005-6	journal	May 2014
Intron-Based Single Transcript Unit CRISPR Systems for Plant Genome Editing Zhong, Zhaohui; Liu, Shishi; Liu, Xiaopei Rice, Vol. 13, Issue 1 https://doi.org/10.1186/s12284-020-0369-8	journal	February 2020
A CRISPR/Cas9 toolkit for multiplex genome editing in plants Xing, Hui-Li; Dong, Li; Wang, Zhi-Ping BMC Plant Biology, Vol. 14, Issue 1 https://doi.org/10.1186/s12870-014-0327-y	journal	November 2014
Targeted mutagenesis using the Agrobacterium tumefaciens-mediated CRISPR-Cas9 system in common wheat Zhang, Shujuan; Zhang, Rongzhi; Song, Guoqi BMC Plant Biology, Vol. 18, Issue 1 https://doi.org/10.1186/s12870-018-1496-x	journal	November 2018
CRISPR/Cas9-mediated genome editing induces gene knockdown by altering the pre-mRNA splicing in mice Tang, Ji-Xin; Chen, Dafeng; Deng, Shou-Long BMC Biotechnology, Vol. 18, Issue 1 https://doi.org/10.1186/s12896-018-0472-8	journal	October 2018
CRISPR-Cas9 mediated targeted disruption of FAD2–2 microsomal omega-6 desaturase in soybean (Glycine max.L) al Amin, Noor; Ahmad, Naveed; Wu, Nan BMC Biotechnology, Vol. 19, Issue 1 https://doi.org/10.1186/s12896-019-0501-2	journal	January 2019
A highly efficient ligation-independent cloning system for CRISPR/Cas9 based genome editing in plants Khan, Aftab A.; El-Sayed, Ashraf; Akbar, Asma Plant Methods, Vol. 13, Issue 1 https://doi.org/10.1186/s13007-017-0236-9	journal	October 2017
Optimization of CRISPR/Cas9 genome editing in cotton by improved sgRNA expression Long, Lu; Guo, Dan-Dan; Gao, Wei Plant Methods, Vol. 14, Issue 1 https://doi.org/10.1186/s13007-018-0353-0	journal	October 2018
An optimised CRISPR/Cas9 protocol to create targeted mutations in homoeologous genes and an efficient genotyping protocol to identify edited events in wheat Cui, Xiucheng; Balcerzak, Margaret; Schernthaner, Johann Plant Methods, Vol. 15, Issue 1 https://doi.org/10.1186/s13007-019-0500-2	journal	October 2019
Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation Wang, Zhi-Ping; Xing, Hui-Li; Dong, Li Genome Biology, Vol. 16, Issue 1 https://doi.org/10.1186/s13059-015-0715-0	journal	July 2015
Integrated metabolomics and phytochemical genomics approaches for studies on rice Okazaki, Yozo; Saito, Kazuki GigaScience, Vol. 5, Issue 1 https://doi.org/10.1186/s13742-016-0116-7	journal	March 2016
Making designer mutants in model organisms Peng, Ying; Clark, Karl J.; Campbell, Jarryd M. Development, Vol. 141, Issue 21 https://doi.org/10.1242/dev.102186	journal	November 2014
Epigenetic Alterations at Genomic Loci Modified by Gene Targeting in Arabidopsis thaliana Lieberman-Lazarovich, Michal; Melamed-Bessudo, Cathy; de Pater, Sylvia PLoS ONE, Vol. 8, Issue 12 https://doi.org/10.1371/journal.pone.0085383	journal	December 2013
sgRNAcas9: A Software Package for Designing CRISPR sgRNA and Evaluating Potential Off-Target Cleavage Sites Xie, Shengsong; Shen, Bin; Zhang, Chaobao PLoS ONE, Vol. 9, Issue 6 https://doi.org/10.1371/journal.pone.0100448	journal	June 2014
Pervasive Genotypic Mosaicism in Founder Mice Derived from Genome Editing through Pronuclear Injection Oliver, Daniel; Yuan, Shuiqiao; McSwiggin, Hayden PLOS ONE, Vol. 10, Issue 6 https://doi.org/10.1371/journal.pone.0129457	journal	June 2015
Enhanced Rice Blast Resistance by CRISPR/Cas9-Targeted Mutagenesis of the ERF Transcription Factor Gene OsERF922 Wang, Fujun; Wang, Chunlian; Liu, Piqing PLOS ONE, Vol. 11, Issue 4 https://doi.org/10.1371/journal.pone.0154027	journal	April 2016
A donor-specific QTL, exhibiting allelic variation for leaf sheath hairiness in a nested association mapping population, is located on barley chromosome 4H Saade, Stephanie; Kutlu, Burcu; Draba, Vera PLOS ONE, Vol. 12, Issue 12 https://doi.org/10.1371/journal.pone.0189446	journal	December 2017
A simple test for the cleavage activity of customized endonucleases in plants Budhagatapalli, Nagaveni; Schedel, Sindy; Gurushidze, Maia RWTH Aachen University https://doi.org/10.18154/rwth-2016-02558	text	January 2016
CRISPR/Cas9 Trends and Future Scheme for Genome Engineering in Plants Paudel, Niroj; Park, Suhyong; Jang, Yoonah Preprints.org https://doi.org/10.20944/preprints202103.0277.v1	journal	March 2021
Biotechnological strategies and tools for Plum pox virus resistance: trans-, intra-, cis-genesis, and beyond Ilardi, Vincenza; Tavazza, Mario Frontiers in Plant Science, Vol. 6 https://doi.org/10.3389/fpls.2015.00379	journal	June 2015
Application of genomics-assisted breeding for generation of climate resilient crops: progress and prospects Kole, Chittaranjan; Muthamilarasan, Mehanathan; Henry, Robert Frontiers in Plant Science, Vol. 6 https://doi.org/10.3389/fpls.2015.00563	journal	August 2015
DNA damage and repair in plants – from models to crops Manova, Vasilissa; Gruszka, Damian Frontiers in Plant Science, Vol. 6 https://doi.org/10.3389/fpls.2015.00885	journal	October 2015
Targeting Non-Coding RNAs in Plants with the CRISPR-Cas Technology is a Challenge yet Worth Accepting Basak, Jolly; Nithin, Chandran Frontiers in Plant Science, Vol. 6 https://doi.org/10.3389/fpls.2015.01001	journal	November 2015
Expanding Omics Resources for Improvement of Soybean Seed Composition Traits Chaudhary, Juhi; Patil, Gunvant B.; Sonah, Humira Frontiers in Plant Science, Vol. 6 https://doi.org/10.3389/fpls.2015.01021	journal	November 2015
Reassessment of the Four Yield-related Genes Gn1a, DEP1, GS3, and IPA1 in Rice Using a CRISPR/Cas9 System Li, Meiru; Li, Xiaoxia; Zhou, Zejiao Frontiers in Plant Science, Vol. 7 https://doi.org/10.3389/fpls.2016.00377	journal	March 2016
Recent Advances in Genome Editing Using CRISPR/Cas9 Ding, Yuduan; Li, Hong; Chen, Ling-Ling Frontiers in Plant Science, Vol. 7 https://doi.org/10.3389/fpls.2016.00703	journal	May 2016
Efficient Inactivation of Symbiotic Nitrogen Fixation Related Genes in Lotus japonicus Using CRISPR-Cas9 Wang, Longxiang; Wang, Longlong; Tan, Qian Frontiers in Plant Science, Vol. 7 https://doi.org/10.3389/fpls.2016.01333	journal	August 2016
Recent Developments in Systems Biology and Metabolic Engineering of Plant–Microbe Interactions Kumar, Vishal; Baweja, Mehak; Singh, Puneet K. Frontiers in Plant Science, Vol. 7 https://doi.org/10.3389/fpls.2016.01421	journal	September 2016
Genome Editing in Sugarcane: Challenges Ahead Mohan, Chakravarthi Frontiers in Plant Science, Vol. 7 https://doi.org/10.3389/fpls.2016.01542	journal	October 2016
Corrigendum: Building a Genetic Manipulation Tool Box for Orchid Biology: Identification of Constitutive Promoters and Application of CRISPR/Cas9 in the Orchid, Dendrobium officinale Kui, Ling Frontiers in Plant Science, Vol. 8 https://doi.org/10.3389/fpls.2017.00664	journal	January 2017
Gene Editing and Crop Improvement Using CRISPR-Cas9 System Arora, Leena; Narula, Alka Frontiers in Plant Science, Vol. 8 https://doi.org/10.3389/fpls.2017.01932	journal	November 2017
Lycopene Is Enriched in Tomato Fruit by CRISPR/Cas9-Mediated Multiplex Genome Editing Li, Xindi; Wang, Yanning; Chen, Sha Frontiers in Plant Science, Vol. 9 https://doi.org/10.3389/fpls.2018.00559	journal	April 2018
Application of CRISPR/Cas9 Genome Editing Technology for the Improvement of Crops Cultivated in Tropical Climates: Recent Progress, Prospects, and Challenges Haque, Effi; Taniguchi, Hiroaki; Hassan, Md. Mahmudul Frontiers in Plant Science, Vol. 9 https://doi.org/10.3389/fpls.2018.00617	journal	May 2018
Non-TAL Effectors From Xanthomonas oryzae pv. oryzae Suppress Peptidoglycan-Triggered MAPK Activation in Rice Long, Juying; Song, Congfeng; Yan, Fang Frontiers in Plant Science, Vol. 9 https://doi.org/10.3389/fpls.2018.01857	journal	December 2018
Harnessing Genome Editing Techniques to Engineer Disease Resistance in Plants Mushtaq, Muntazir; Sakina, Aafreen; Wani, Shabir Hussain Frontiers in Plant Science, Vol. 10 https://doi.org/10.3389/fpls.2019.00550	journal	May 2019
Prospects of Understanding the Molecular Biology of Disease Resistance in Rice Singh, Pankaj; Nag, Akshay; Arya, Preeti International Journal of Molecular Sciences, Vol. 19, Issue 4 https://doi.org/10.3390/ijms19041141	journal	April 2018
CRISPR/Cas9-Mediated Multiplex Genome Editing of the BnWRKY11 and BnWRKY70 Genes in Brassica napus L. Sun, Qinfu; Lin, Li; Liu, Dongxiao International Journal of Molecular Sciences, Vol. 19, Issue 9 https://doi.org/10.3390/ijms19092716	journal	September 2018
Highly Efficient Targeted Gene Editing in Upland Cotton Using the CRISPR/Cas9 System Zhu, Shouhong; Yu, Xiuli; Li, Yanjun International Journal of Molecular Sciences, Vol. 19, Issue 10 https://doi.org/10.3390/ijms19103000	journal	October 2018
Fertility of Pedicellate Spikelets in Sorghum Is Controlled by a Jasmonic Acid Regulatory Module Gladman, Nicholas; Jiao, Yinping; Lee, Young Koung International Journal of Molecular Sciences, Vol. 20, Issue 19 https://doi.org/10.3390/ijms20194951	journal	October 2019
Tipping Points in Seaweed Genetic Engineering: Scaling Up Opportunities in the Next Decade Lin, Hanzhi; Qin, Song Marine Drugs, Vol. 12, Issue 5 https://doi.org/10.3390/md12053025	journal	May 2014
Allelic Variants of CRISPR/Cas9 Induced Mutation in an Inositol Trisphosphate 5/6 Kinase Gene Manifest Different Phenotypes in Barley No authors listed Plants, Vol. 9, Issue 2 https://doi.org/10.3390/plants9020195	journal	February 2020
CRISPR system for genome engineering: the application for autophagy study Cui, Jianzhou; Chew, Shirley Jia Li; Shi, Yin BMB Reports, Vol. 50, Issue 5 https://doi.org/10.5483/bmbrep.2017.50.5.044	journal	May 2017
Additional file 2 of Application of CRISPR/Cas9 technology in wild apple (Malus sieverii) for paired sites gene editing Zhang, Yan; Zhou, Ping; Bozorov, Tohir A. figshare https://doi.org/10.6084/m9.figshare.15020573.v1	dataset	January 2021
Additional file 3 of Application of CRISPR/Cas9 technology in wild apple (Malus sieverii) for paired sites gene editing Zhang, Yan; Zhou, Ping; Bozorov, Tohir A. figshare https://doi.org/10.6084/m9.figshare.15020576	dataset	January 2021
Additional file 4 of Application of CRISPR/Cas9 technology in wild apple (Malus sieverii) for paired sites gene editing Zhang, Yan; Zhou, Ping; Bozorov, Tohir A. figshare https://doi.org/10.6084/m9.figshare.15020579	dataset	January 2021
Additional file 5 of Application of CRISPR/Cas9 technology in wild apple (Malus sieverii) for paired sites gene editing Zhang, Yan; Zhou, Ping; Bozorov, Tohir A. figshare https://doi.org/10.6084/m9.figshare.15020582.v1	dataset	January 2021
Improvement of the CRISPR-Cpf1 system with ribozyme-processed crRNA Gao, Zongliang; Herrera-Carrillo, Elena; Berkhout, Ben Taylor & Francis https://doi.org/10.6084/m9.figshare.7379300.v1	text	January 2018

Similar Records

Efficient CRISPR/Cas9-Mediated Gene Editing in Arabidopsis thaliana and Inheritance of Modified Genes in the T2 and T3 Generations

Journal Article · Tue Jun 10 20:00:00 EDT 2014 · PLoS ONE · OSTI ID:1904806

A gene-within-a-gene Cas9/sgRNA hybrid construct enables gene editing and gene replacement strategies in Chlamydomonas reinhardtii

Journal Article · Mon Apr 24 20:00:00 EDT 2017 · Algal Research · OSTI ID:1533503

Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs

Journal Article · Wed May 08 20:00:00 EDT 2019 · Nature Communications · OSTI ID:1603511

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
assembly cloning
synthetic biology

Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice

Citation Formats

References (23)

Cited By (67)

Similar Records

Related Subjects