Generation of multi-gene knockout rabbits using the Cas9/gRNA system

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• 作者：Quanmei Yan (1)
  Quanjun Zhang (1)
  Huaqiang Yang (1)
  Qingjian Zou (1)
  Chengcheng Tang (2)
  Nana Fan (1)
  Liangxue Lai (1) (2)
  
  1. Key Laboratory of Regenerative Biology ; South China Institute for Stem Cell Biology and Regenerative Medicine ; Guangzhou Institutes of Biomedicine and Health ; Chinese Academy of Sciences ; Guangzhou ; 510530 ; China
  2. College of Animal Science ; Jilin University ; Changchun ; 130062 ; China
  
• 关键词：Cas9/gRNA system ; Rabbits ; Multiple ; gene knockout
• 刊名：Cell Regeneration
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：3
• 期：1
• 全文大小：764 KB
• 参考文献：1. Li, XJ, Li, W (2012) Beyond mice: genetically modifying larger animals to model human diseases. J Genet Genomics 39: pp. 237-238 CrossRef
  2. Seok, J, Warren, HS, Cuenca, AG, Mindrinos, MN, Baker, HV, Xu, W, Richards, DR, McDonald-Smith, GP, Gao, H, Hennessy, L, Finnerty, CC, Lopez, CM, Honari, S, Moore, EE, Minei, JP, Cuschieri, J, Bankey, PE, Johnson, JL, Sperry, J, Nathens, AB, Billiar, TR, West, MA, Jeschke, MG, Klein, MB, Gamelli, RL, Gibran, NS, Brownstein, BH, Miller-Graziano, C, Calvano, SE, Mason, PH (2013) Genomic responses in mouse models poorly mimic human inflammatory diseases. Proc Natl Acad Sci U S A 110: pp. 3507-3512 CrossRef
  3. Yu Wang, NF, Jun, S, Juan, Z, Xiaogang, G, Weihua, T, Quanjun, Z, Fenggong, C, Li Li, PNN, Jon, F, Esteban, MA, Liangxue, L (2014) Generation of knockout rabbits using transcription activator-like effector nucleases. Cell Regeneration 3: pp. 9 CrossRef
  4. Porteus, MH, Carroll, D (2005) Gene targeting using zinc finger nucleases. Nat Biotechnol 23: pp. 967-973 CrossRef
  5. Bogdanove, AJ, Voytas, DF (2011) TAL effectors: customizable proteins for DNA targeting. Science 333: pp. 1843-1846 CrossRef
  6. Zakhartchenko, V, Flisikowska, T, Li, S, Richter, T, Wieland, H, Durkovic, M, Rottmann, O, Kessler, B, Gungor, T, Brem, G, Kind, A, Wolf, E, Schnieke, A (2011) Cell-mediated transgenesis in rabbits: chimeric and nuclear transfer animals. Biol Reprod 84: pp. 229-237 CrossRef
  7. Flisikowska, T, Thorey, IS, Offner, S, Ros, F, Lifke, V, Zeitler, B, Rottmann, O, Vincent, A, Zhang, L, Jenkins, S, Niersbach, H, Kind, AJ, Gregory, PD, Schnieke, AE, Platzer, J (2011) Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases. PLoS One 6: pp. e21045 CrossRef
  8. Song, J, Zhong, J, Guo, X, Chen, Y, Zou, Q, Huang, J, Li, X, Zhang, Q, Jiang, Z, Tang, C, Yang, H, Liu, T, Li, P, Pei, D, Lai, L (2013) Generation of RAG 1- and 2-deficient rabbits by embryo microinjection of TALENs. Cell Res 23: pp. 1059-1062 CrossRef
  9. Bhaya, D, Davison, M, Barrangou, R (2011) CRISPR-Cas systems in bacteria and archaea: versatile small RNAs for adaptive defense and regulation. Annu Rev Genet 45: pp. 273-297 CrossRef
  10. Wiedenheft, B, Sternberg, SH, Doudna, JA (2012) RNA-guided genetic silencing systems in bacteria and archaea. Nature 482: pp. 331-338 CrossRef
  11. Gratz, SJ, Cummings, AM, Nguyen, JN, Hamm, DC, Donohue, LK, Harrison, MM, Wildonger, J, O'Connor-Giles, KM (2013) Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Genetics 194: pp. 1029-1035 CrossRef
  12. Bassett, AR, Tibbit, C, Ponting, CP, Liu, JL (2013) Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system. Cell Rep 4: pp. 220-228 CrossRef
  13. Friedland, AE, Tzur, YB, Esvelt, KM, Colaiacovo, MP, Church, GM, Calarco, JA (2013) Heritable genome editing in C. elegans via a CRISPR-Cas9 system. Nat Methods 10: pp. 741-743 CrossRef
  14. Xie, K, Yang, Y (2013) RNA-guided genome editing in plants using a CRISPR-Cas system. Mol Plant 6: pp. 1975-1983 CrossRef
  15. Chang, N, Sun, C, Gao, L, Zhu, D, Xu, X, Zhu, X, Xiong, JW, Xi, JJ (2013) Genome editing with RNA-guided Cas9 nuclease in zebrafish embryos. Cell Res 23: pp. 465-472 CrossRef
  16. Jao, LE, Wente, SR, Chen, W (2013) Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. Proc Natl Acad Sci U S A 110: pp. 13904-13909 CrossRef
  17. Hwang, WY, Fu, Y, Reyon, D, Maeder, ML, Kaini, P, Sander, JD, Joung, JK, Peterson, RT, Yeh, JR (2013) Heritable and Precise Zebrafish Genome Editing Using a CRISPR-Cas System. PLoS One 8: pp. e68708 CrossRef
  18. Wang, H, Yang, H, Shivalila, CS, Dawlaty, MM, Cheng, AW, Zhang, F, Jaenisch, R (2013) One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering. Cell 153: pp. 910-918 CrossRef
  19. Li, D, Qiu, Z, Shao, Y, Chen, Y, Guan, Y, Liu, M, Li, Y, Gao, N, Wang, L, Lu, X, Zhao, Y (2013) Heritable gene targeting in the mouse and rat using a CRISPR-Cas system. Nat Biotechnol 31: pp. 681-683 CrossRef
  20. Li, W, Teng, F, Li, T, Zhou, Q (2013) Simultaneous generation and germline transmission of multiple gene mutations in rat using CRISPR-Cas systems. Nat Biotechnol 31: pp. 684-686 CrossRef
  21. Tan, WF, Carlson, DF, Lancto, CA, Garbe, JR, Webster, DA, Hackett, PB, Fahrenkrug, SC (2013) Efficient nonmeiotic allele introgression in livestock using custom endonucleases. Proc Natl Acad Sci U S A 110: pp. 16526-16531 CrossRef
  22. Ding, Q, Regan, SN, Xia, Y, Oostrom, LA, Cowan, CA, Musunuru, K (2013) Enhanced Efficiency of Human Pluripotent Stem Cell Genome Editing through Replacing TALENs with CRISPRs. Cell Stem Cell 12: pp. 393-394 CrossRef
  23. Mali, P, Yang, L, Esvelt, KM, Aach, J, Guell, M, DiCarlo, JE, Norville, JE, Church, GM (2013) RNA-guided human genome engineering via Cas9. Science 339: pp. 823-826 CrossRef
  24. Niu, Y, Shen, B, Cui, Y, Chen, Y, Wang, J, Wang, L, Kang, Y, Zhao, X, Si, W, Li, W, Xiang, AP, Zhou, J, Guo, X, Bi, Y, Si, C, Hu, B, Dong, G, Wang, H, Zhou, Z, Li, T, Tan, T, Pu, X, Wang, F, Ji, S, Zhou, Q, Huang, X, Ji, W, Sha, J (2014) Generation of Gene-Modified Cynomolgus Monkey via Cas9/RNA-Mediated Gene Targeting in One-Cell Embryos. Cell 156: pp. 836-843 CrossRef
  25. Yang, D, Xu, J, Zhu, T, Fan, J, Lai, L, Zhang, J, Chen, YE (2014) Effective gene targeting in rabbits using RNA-guided Cas9 nucleases. J Mol Cell Biol 6: pp. 97-99 CrossRef
  26. Jinek, M, Chylinski, K, Fonfara, I, Hauer, M, Doudna, JA, Charpentier, E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337: pp. 816-821 CrossRef
  27. Cong, L, Ran, FA, Cox, D, Lin, S, Barretto, R, Habib, N, Hsu, PD, Wu, X, Jiang, W, Marraffini, LA, Zhang, F (2013) Multiplex genome engineering using CRISPR/Cas systems. Science 339: pp. 819-823 CrossRef
  28. Zhang, X, Abreu, JG, Yokota, C, MacDonald, BT, Singh, S, Coburn, KL, Cheong, SM, Zhang, MM, Ye, QZ, Hang, HC, Steen, H, He, X (2012) Tiki1 is required for head formation via Wnt cleavage-oxidation and inactivation. Cell 149: pp. 1565-1577 CrossRef
  29. Pearson, T, Shultz, LD, Miller, D, King, M, Laning, J, Fodor, W, Cuthbert, A, Burzenski, L, Gott, B, Lyons, B, Foreman, O, Rossini, AA, Greiner, DL (2008) Non-obese diabetic-recombination activating gene-1 (NOD-Rag1 null) interleukin (IL)-2 receptor common gamma chain (IL2r gamma null) null mice: a radioresistant model for human lymphohaematopoietic engraftment. Clin Exp Immunol 154: pp. 270-284 CrossRef
  30. Fu, Y, Sander, JD, Reyon, D, Cascio, VM, Joung, JK (2014) Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat Biotechnol 32: pp. 279-284 CrossRef
  31. Ran, FA, Hsu, PD, Lin, CY, Gootenberg, JS, Konermann, S, Trevino, AE, Scott, DA, Inoue, A, Matoba, S, Zhang, Y, Zhang, F (2013) Double Nicking by RNA-Guided CRISPR Cas9 for Enhanced Genome Editing Specificity (vol 154, pg 1380, 2013). Cell 155: pp. 479-480 CrossRef
  32. Fu, Y, Foden, JA, Khayter, C, Maeder, ML, Reyon, D, Joung, JK, Sander, JD (2013) High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Nat Biotechnol 31: pp. 822-826 CrossRef
  33. Tian, J, Song, J, Li, H, Yang, D, Li, X, Ouyang, H, Lai, L (2012) Effect of donor cell type on nuclear remodelling in rabbit somatic cell nuclear transfer embryos. Reprod Domest Anim 47: pp. 544-552 CrossRef
  
• 刊物主题：Cell Biology; Stem Cells; Biomedicine general;
• 出版者：BioMed Central
• ISSN：2045-9769

文摘

The prokaryotic clustered regularly interspaced short palindromic repeat (CRISPR)-associated system (Cas) is a simple, robust and efficient technique for gene targeting in model organisms such as zebrafish, mice and rats. In this report, we applied CRISPR technology to rabbits by microinjection of Cas9 mRNA and guided RNA (gRNA) into the cytoplasm of pronuclear-stage embryos. We achieved biallelic gene knockout (KO) rabbits by injection of 1 gene (IL2rg) or 2 gene (IL2rg and RAG1) Cas9 mRNA and gRNA with an efficiency of 100%. We also tested the efficiency of multiple gene KOs in early rabbit embryos and found that the efficiency of simultaneous gene mutation on target sites is as high as 100% for 3 genes (IL2rg, RAG1 and RAG2) and 33.3% for 5 genes (IL2rg, RAG1, RAG2, TIKI1 and ALB). Our results demonstrate that the Cas9/gRNA system is a highly efficient and fast tool not only for single-gene editing but also for multi-gene editing in rabbits.