遗传 ›› 2015, Vol. 37 ›› Issue (10): 974-982.doi: 10.16288/j.yczz.15-109
璩良, 李华善, 姜运涵, 董春升
收稿日期:2015-03-16
出版日期:2015-10-20
发布日期:2015-10-20
通讯作者:
董春升，副教授，研究方向：免疫学。 E-mail：chunshengdong@suda.edu.cn
作者简介:璩良，本科生，专业方向：生物技术(免疫工程)。E-mail: quliangsuda@163.com
基金资助:Liang Qu， Huashan Li， Yunhan Jiang， Chunsheng Dong
Received:2015-03-16
Online:2015-10-20
Published:2015-10-20
摘要： CRISPR/Cas系统是广泛存在于细菌和古生菌中的适应性免疫系统，用来抵抗外来病毒或质粒的入侵。近几年，由Ⅱ型CRISPR/Cas适应性免疫系统改造而来的CRISPR/ Cas9基因组编辑技术蓬勃发展，被广泛地应用于生命科学研究的各个领域，并取得了革命性的变化。文章主要综述了CRISPR/Cas9基因组编辑技术的起源与发展及在生命科学各研究领域的应用，重点介绍了该系统在人类疾病基因治疗方面的最新应用及脱靶效应，以期为相关领域的科研人员提供参考。
璩良, 李华善, 姜运涵, 董春升. CRISPR/Cas9系统的分子机制及其在人类疾病基因治疗中的应用[J]. 遗传, 2015, 37(10): 974-982.
Liang Qu， Huashan Li， Yunhan Jiang， Chunsheng Dong. The molecular mechanism of CRISPR/Cas9 system and its application in gene therapy of human diseases[J]. HEREDITAS(Beijing), 2015, 37(10): 974-982.
| [1] Keeney S, Giroux CN, Kleckner N. Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell, 1997, 88(3): 375–384. [2] Szostak JW, Orr-Weaver TL, Rothstein RJ, Stahl FW. The double-strand-break repair model for recombination. Cell, 1983, 33(1): 25–35. [3] Miller JC, Holmes MC, Wang JB, Guschin DY, Lee YL, Rupniewski I, Beausejour CM, Waite AJ, Wang NS, Kim KA, Gregory PD, Pabo CO, Rebar EJ. An improved zinc-finger nuclease architecture for highly specific genome editing. Nat Biotechnol, 2007, 25(7): 778–785. [4] Urnov FD, Miller JC, Lee YL, Beausejour CM, Rock JM, Augustus S, Jamieson AC, Porteus MH, Gregory PD, Holmes MC. Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature, 2005, 435(7042): 646–651. [5] Boch J, Scholze H, Schornack S, Landgraf A, Hahn S, Kay S, Lahaye T, Nickstadt A, Bonas U. Breaking the code of DNA binding specificity of TAL-type III effectors. Science, 2009, 326(5959): 1509–1512. [6] Christian M, Cermak T, Doyle EL, Schmidt C, Zhang F, Hummel A, Bogdanove AJ, Voytas DF. Targeting DNA double-strand breaks with TAL effector nucleases. Genetics, 186(2): 757–761. [7] Miller JC, Tan SY, Qiao GJ, Barlow KA, Wang JB, Xia DF, Meng XD, Paschon DE, Leung E, Hinkley SJ, Dulay GP, Hua KL, Ankoudinova I, Cost GJ, Urnov FD, Zhang HS, Holmes MC, Zhang L, Gregory PD, Rebar EJ. A TALE nuclease architecture for efficient genome editing. Nat Biotechnol, 2011, 29(2): 143–148. [8] Cong L, Ran FA, Cox D, Lin SL, Barretto R, Habib N, Hsu PD, Wu XB, Jiang WY, Marraffini LA, Zhang F. Multiplex genome engineering using CRISPR/Cas systems. Science, 2013, 339(6121): 819–823. [9] Jiang WY, Bikard D, Cox D, Zhang F Marraffini LA. RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol, 2013, 31(3): 233–239. [10] Mali P, Yang LH, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE, Church GM. RNA-guided human genome engineering via Cas9. Science, 2013, 339(6121): 823–826. [11] Wang HY, Yang H, Shivalila CS, Dawlaty MM, Cheng AW, Zhang F, Jaenisch R. One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell, 2013, 153(4): 910–918. [12] Sroubek J, Krishnan Y McDonald TV. Sequence and structure-specific elements of HERG mRNA determine channel synthesis and trafficking efficiency. FASEB J, 2013, 27(8): 3039–3053. [13] Wiedenheft B, van Duijn E, Bultema JB, Waghmare SP, Zhou KH, Barendregt A, Westphal W, Heck AJR, Boekema EJ. Dickman MJ, Doudna JA. RNA-guided complex from a bacterial immune system enhances target recognition through seed sequence interactions. Proc Natl Acad Sci USA, 2011, 108(25): 10092–10097. [14] Hale CR, Zhao P, Olson S, Duff MO, Graveley BR, Wells L, Terns RM, Terns MP. RNA-guided RNA cleavage by a CRISPR RNA-Cas protein complex. Cell, 2009, 139(5): 945–956. [15] Marraffini LA, Sontheimer EJ. CRISPR interference limits horizontal gene transfer in staphylococci by targeting DNA. Science, 2008, 322(5909): 1843–1845. [16] Deltcheva E, Chylinski K, Sharma CM, Gonzales K, Chao YJ, Pirzada ZA, Eckert MR, Vogel J, Charpentier E. CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. Nature, 2011, 471(7340): 602–607. [17] Garneau JE, Dupuis Mè, Villion M, Romero DA, Barrangou R, Boyaval P, Fremaux C, Horvath P, Magadán AH, Moineau S. The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA. Nature, 2010, 468(7320): 67–71. [18] Heler R, Marraffini LA, Bikard D. Adapting to new threats: the generation of memory by CRISPR-Cas immune systems. Mol Microbiol, 2014, 93(1): 1–9. [19] Jansen R, van Embden JDA, Gaastra W, Schouls LM. Identification of genes that are associated with DNA repeats in prokaryotes. Mol Microbiol, 2002, 43(6): 1565–1575. [20] Mojica FJ, Díez-Villase?or C, Soria E, Juez G. Biological significance of a family of regularly spaced repeats in the genomes of Archaea, Bacteria and mitochondria. Mol Microbiol, 2014, 36(1): 244–246. [21] 张智辉, 董少忠, 寸韡. 基因组定点编辑技术的研究进展. 生命科学, 2013, 25(7): 735–742. [22] 李铁民, 杜波. CRISPR-Cas系统与细菌和噬菌体的共进化. 遗传, 2011, 33(3): 213–218. [23] 王菲, 罗恩杰. CRISPR及其在原核生物防御系统中的作用. 热带医学杂志, 2008, 8(10): 1104–1106. [24] Chylinski K, Le Rhun A, Charpentier E. The tracrRNA and Cas9 families of type II CRISPR-Cas immunity systems. RNA Biol, 2013, 10(5): 726–737. [25] Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science, 2012, 337(6096): 816–821. [26] Hwang WY, Fu YF, Reyon D, Maeder ML, Tsai SQ, Sander JD, Peterson RT, Yeh JR, Joung JK. Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol, 2013, 31(3): 227–229. [27] Chylinski K, Makarova KS, Charpentier E, Koonin EV. Classification and evolution of type II CRISPR-Cas systems. Nucleic Acids Res, 2014, 42(10): 6091–6105. [28] Liu YY, Ma SY, Wang XG, Chang JS, Gao J, Shi R, Zhang JD, Lu W, Zhao P, Xia QY. Highly efficient multiplex targeted mutagenesis and genomic structure variation in Bombyx mori cells using CRISPR/Cas9. Insect Biochem Mol Biol, 2014, 49: 35–42. [29] Auer TO, Del Bene F. CRISPR/Cas9 and TALEN-mediated knock-in approaches in zebrafish. Methods, 2014, 69(2): 142–150. [30] Ran FA, Hsu PD, Lin CY, Gootenberg JS, Konermann S, Trevino AE, Scott DA, Inoue A, Matoba S, Zhang Y, Zhang F. Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell, 2013, 154(6): 1380–1389. [31] Yin H, Xue W, Chen SD, Bogorad RL, Benedetti E, Grompe M, Koteliansky V, Sharp PA, Jacks T, Anderson DG. Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype. Nat Biotechnol, 2014, 32(6): 551–553. [32] Ma YW, Ma J, Zhang X, Chen W, Yu L, Lu YD, Bai L, Shen B, Huang XX, Zhang LF. Generation of eGFP and Cre knockin rats by CRISPR/Cas9. FEBS J, 2014, 281(17): 3779–3790. [33] Zhang C, Xiao B, Jiang YY, Zhao YH, Li ZK, Gao H, Ling Y, Wei J, Li SN, Lu MK, Su XZ, Cui HT, Yuan J. Efficient editing of malaria parasite genome using the CRISPR/Cas9 system. MBio, 2014, 5(4): e01414-14. [34] Long CZ, McAnally JR, Shelton JM, Mireault AA, Bassel-Duby R, Olson EN. Prevention of muscular dystrophy in mice by CRISPR/Cas9-mediated editing of germline DNA. Science, 2014, 345(6201): 1184–1188. [35] Xie F, Ye L, Chang JC, Beyer AI, Wang JM, Muench MO, Kan YW. Seamless gene correction of β-thalassemia mutations in patient-specific iPSCs using CRISPR/Cas9 and piggyBac. Genome Res, 2014, 24(9): 1526–1533. [36] Wu YX, Liang D, Wang YH, Bai MZ, Tang W, Bao SM, Yan ZQ, Li DS, Li JS. Correction of a genetic disease in mouse via use of CRISPR-Cas9. Cell Stem Cell, 2013, 13(6): 659–662. [37] Manjunath N, Yi GH, DangY, Shankar P. Newer gene editing technologies toward HIV gene therapy. Viruses, 2013, 5(11): 2748–2766. [38] Hu WH, Kaminski R, Yang F, Zhang YG, Cosentino L, Li F, Luo B, Alvarez-Carbonell D, Garcia-Mesa Y, Karn J, Mo XM, Khalili K. RNA-directed gene editing specifically eradicates latent and prevents new HIV-1 infection. Proc Natl Acad Sci USA, 2014, 111(31): 11461–11466. [39] Dong CS, Qu L, Wang HY, Wei L, Dong YS, Xiong SD. Targeting hepatitis B virus cccDNA by CRISPR/Cas9 nuclease efficiently inhibits viral replication. Antiviral Res, 2015, 118: 110–117. [40] Kennedy EM, Bassit LC, Mueller H, Kornepati AVR, Bogerd HP, Nie T, Chatterjee P, Javanbakht H, Schinazi RF, Cullen BR. Suppression of hepatitis B virus DNA accumulation in chronically infected cells using a bacterial CRISPR/Cas RNA-guided DNA endonuclease. Virology, 2015, 476: 196–205. [41] Lin SR, Yang HC, Kuo YT, Liu CJ, Yang TY, Sung KC, Lin YY, Wang HY, Wang CC, Shen YC, Wu FY, Kao JH, Chen DS, Chen PJ. The CRISPR/Cas9 system facilitates clearance of the intrahepatic HBV templates in vivo. Mol Ther Nucleic Acids, 2014, 3(8): e186. [42] Seeger C, Sohn JA. Targeting hepatitis B virus with CRISPR/Cas9. Mol Ther Nucleic Acids, 2014, 3(12): e216. [43] Zhen S, Hua L, Liu YH, Gao LC, Fu J, Wan DY, Dong LH, Song HF, Gao X. Harnessing the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated Cas9 system to disrupt the hepatitis B virus. Gene Ther, 2015, 22(5): 404–412. [44] Suenaga T, Kohyama M, Hirayasu K, Arase H. Engineering large viral DNA genomes using the CRISPR-Cas9 system. Microbiol Immunol, 2014, 58(9): 513–522. [45] Torres R, Martin MC, Garcia A, Cigudosa JC, Ramirez JC, Rodriguez-Perales S. Engineering human tumour- associated chromosomal translocations with the RNA-guided CRISPR-Cas9 system. Nat Commun, 2014, 5: 3964. [46] Xue W, Chen SD, Yin H, Tammela T, Papagiannakopoulos T, Joshi NS, Cai WX, Yang G, Bronson R, Crowley DG, Zhang F, Anderson DG, Sharp PA, Jacks T. CRISPR-mediated direct mutation of cancer genes in the mouse liver. Nature, 2014, 514(7522): 380–384. [47] Platt RJ, Chen SD, Zhou Y, Yim MJ, Swiech L, Kempton HR, Dahlman JE, Parnas O, Eisenhaure TM, Jovanovic M, Graham DB, Jhunjhunwala S, Heidenreich M, Xavier RJ, Langer R, Anderson DG, Hacohen N, Regev A, Feng GP, Sharp PA, Zhang F. CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell, 2014, 159(2): 440–455. [48] Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. Genome-scale CRISPR-Cas9 knockout screening in human cells. Science, 2014, 343(6166): 84–87. [49] Wang T, Wei JJ, Sabatini DM, Lander ES. Genetic screens in human cells using the CRISPR-Cas9 system. Science, 2014, 343(6166): 80–84. [50] Zhou YX, Zhu SY, Cai CZ, Yuan PF, Li CM, Huang YY, Wei WS. High-throughput screening of a CRISPR/Cas9 library for functional genomics in human cells. Nature, 2014, 509(7501): 487–491. [51] Cheng AW, Wang HY, Yang H, Shi LY, Katz Y, Theunissen TW, Rangarajan S, Shivalila CS, Dadon DB, Jaenisch R. Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system. Cell Res, 2013, 23(10): 1163–1171. [52] Gilbert LA, Larson MH, Morsut L, Liu ZR, Brar GA, Torres SE, Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA, Weissman JS, Qi LS. CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes. Cell, 2013, 154(2): 442–451. [53] Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim WA. Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell, 2013, 152(5): 1173–1183. [54] Chen BH, Gilbert LA, Cimini BA, Schnitzbauer J, Zhang W, Li GW, Park J, Blackburn EH, Weissman JS, Qi LS, Huang B. Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system. Cell, 2013, 155(7): 1479–1491. [55] Duan JZ, Lu GQ, Xie Z, Lou ML, Luo J, Guo L, Zhang Y. Genome-wide identification of CRISPR/Cas9 off-targets in human genome. Cell Res, 2014, 24(8): 1009–1012. [56] Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin MJ, Aryee MJ, Joung JK. Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Nat Biotechnol, 2014, 32(6): 569–576. [57] Guilinger JP, Thompson DB, Liu DR. Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification. Nat Biotechnol, 2014, 32(6): 577–582. [58] Holt N, Wang JB, Kim K, Friedman G, Wang XC, Taupin V, Crooks GM, Kohn DB, Gregory PD, Holmes MC, Cannon PM. Human hematopoietic stem/progenitor cells modified by zinc-finger nucleases targeted to CCR5 control HIV-1 in vivo. Nat Biotechnol, 28(8): 839–847. [59] Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O. Crystal structure of Cas9 in complex with guide RNA and target DNA. Cell, 2014, 156(5): 935–949. [60] Sebastiano V, Maeder ML, Angstman JF, Haddad B, Khayter C, Yeo DT, Goodwin MJ, Hawkins JS, Ramirez CL, Batista LFZ, Artandi SE, Wernig M, Joung JK. In situ genetic correction of the sickle cell anemia mutation in human induced pluripotent stem cells using engineered zinc finger nucleases. Stem Cells, 2011, 29(11): 1717–1726. [61] Yusa K, Rashid ST, Strick-Marchand H, Varela I, Liu PQ, Paschon DE, Miranda E, Ordó?ez A, Hannan NRF, Rouhani FJ, Darche S, Alexander G, Marciniak SJ, Fusaki N, Hasegawa M, Holmes MC, Di Santo JP, Lomas DA, Bradley A, Vallier L. Targeted gene correction of α1-antitrypsin deficiency in induced pluripotent stem cells. Nature, 2011, 478(7369): 391–394. [62] Zou JZ, Mali P, Huang XS, Dowey SN, Cheng LZ. Site-specific gene correction of a point mutation in human iPS cells derived from an adult patient with sickle cell disease. Blood, 2011, 118(17): 4599–4608. [63] Liu YJ, Chen CY, He HX, Wang DS, E LL, Liu ZY, Liu HC. Lentiviral-mediated gene transfer into human adipose-derived stem cells: role of NELL1 versus BMP2 in osteogenesis and adipogenesis in vitro. Acta Biochim Biophys Sin, 2012, 44(10): 856–865. [64] Verma IM, Somia N. Gene therapy-promises, problems and prospects. Nature, 1997, 389(6648): 239–242. [65] Voit RA, McMahon MA, Sawyer SL, Porteus MH. Generation of an HIV resistant T-cell line by targeted "stacking" of restriction factors. Mol Ther, 2013, 21(4): 786–795. [66] Swiech L, Heidenreich M, Banerjee A, Habib N, Li YQ, Trombetta J, Sur M, Zhang F. In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9. Nat Biotechnol, 2015, 33(1): 102–106. [67] Kormann MSD, Hasenpusch G, Aneja MK, Nica G, Flemmer AW, Herber-Jonat S, Huppmann M, Mays LE, Illenyi M, Schams A, Griese M, Bittmann I, Handgretinger R, Hartl D, Rosenecker J, Rudolph C. Expression of therapeutic proteins after delivery of chemically modified mRNA in mice. Nat Biotechnol, 2011, 29(2): 154–157. [68] Zuris JA, Thompson DB, Shu Y, Guilinger JP, Bessen JL, Hu JH, Maeder ML, Joung JK, Chen ZY, Liu DR. Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol, 2015, 33(1): 73–80. |
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