利用生物技术可以对植物基因组进行高效,精准,特异的修饰.锌指核酸酶(Zinc finger nucleases, ZFN),转录激活样效应因子核酸酶(Transcription activator-like effector nucleases, TALEN),成簇规律间隔短回文重复序列(Clustered regularly interspaced short palindromic repeats, CRISPR)/Cas9 (CRISPR-associated 9)是目前基因组编辑技术应用中的关键工程核酸酶.通过产生DNA双链断裂(Double-strand breaks, DSBs)激活植物内源修复途径(包括非同源粘性末端连接和同源重组修复),基因组编辑技术可以实现对靶位点的定点突变,缺失或者基因的插入与替换.基因组编辑已经被广泛地应用到各种植物的基因组修饰中,如拟南芥,水稻,烟草等.本文主要概述了基因组编辑技术在植物基因功能鉴定及作物遗传育种中的应用,并对其未来在作物精准改良中需要完善的相关问题进行了探讨.
Plant genome can be modified via current biotechnology with high specificity and excellent efficiency. Zinc finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system are the key engineered nucleases used in the genome editing. Genome editing techniques enable gene targeted mutagenesis, gene knock-out, gene insertion or replacement at the target sites during the endogenous DNA repair process, including non-homologous end joining (NHEJ) and homologous recombination (HR), triggered by the induction of DNA double-strand break (DSB). Genome editing has been successfully applied in the genome modification of diverse plant species, such as Arabidopsis thaliana, Oryza sativa, and Nicotiana tabacum. In this review, we summarize the application of genome editing in identification of plant gene function and crop breeding. Moreover, we also discuss the improving points of genome editing in crop precision genetic improvement for further study.
[1] Mauricio R. Mapping quantitative trait loci in plants: Uses and caveats for evolutionary biology. Nat Rev Genet , 2001, 2(5): 370-381.
[2] Jorgensen R, Matzke M, Matzke AJM. Plants, RNAi, and the nobel prize. Science , 2006, 314(5803): 1242-1243.
[3] Eamens A, Wang MB, Smith NA, Waterhouse PM. RNA silencing in plants: Yesterday, today, and tomorrow. Plant Physiol , 2008, 147(2): 456-468.
[4] McGinnis KM. RNAi for functional genomics in plants. Brief Funct Genomics , 2010, 9(2): 111-117.
[5] Koncz C, Németh K, Rédei GP, Schell J. T-DNA insertional mutagenesis in Arabidopsis . Plant Mol Biol , 1992, 20(5): 963-976.
[6] Krysan PJ, Young JC, Sussman MR. T-DNA as an insertional mutagen in Arabidopsis . Plant Cell , 1999, 11(12): 2283-2290.
[7] Jeon JS, Lee S, Jung KH, Jun SH, Jeong DH, Lee J, Kim C, Jang S, Lee S, Yang K, Nam J, An K, Han MJ, Sung RJ, Choi HS, Yu JH, Choi JH, Cho SY, Cha SS, Kim SI, An G. T-DNA insertional mutagenesis for functional genomics in rice. Plant J , 2000, 22(6): 561-570.
[8] Jung KH, An G. Functional characterization of rice genes using a gene-indexed T-DNA insertional mutant population. In: Yang YN, ed. Rice Protoc. Humana Press, 2013, 956: 57-67.
[9] Thomas KR, Capecchi MR. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell , 1987, 51(3): 503-512.
[10] Capecchi MR. The new mouse genetics: Altering the genome by gene targeting. Trends Genet , 1989, 5(3): 70-76.
[11] Thomas KR, Capecchi MR. Introduction of homologous DNA sequences into mammalian cells induces mutations in the cognate gene. Nature , 1986, 324(6092): 34-38.
[12] Thomas KR, Folger KR, Capecchi MR. High frequency targeting of genes to specific sites in the mammalian genome. Cell , 1986, 44(3): 419-428.
[13] Doetschman T, Maeda N, Smithies O. Targeted mutation of the Hprt gene in mouse embryonic stem cells. Proc Natl Acad Sci USA , 1988, 85(22): 8583-8587.
[14] Koller BH, Smithies O. Inactivating the β 2 -microglobulin locus in mouse embryonic stem cells by homologous recombination. Proc Natl Acad Sci USA , 1989, 86(22): 8932-8935.
[15] Paszkowski J, Baur M, Bogucki A, Potrykus I. Gene targeting in plants. EMBO J , 1988, 7(13): 4021-4026.
[16] Baur M, Potrykus I, Paszkowski J. Intermolecular homologous recombination in plants. Mol Cell Biol , 1990, 10(2): 492-500.
[17] Lee KY, Lund P, Lowe K, Dunsmuir P. Homologous recombination in plant cells after Agrobacterium -mediated transformation. Plant Cell , 1990, 2(5): 415-425.
[18] Offringa R, de Groot MJA, Haagsman HJ, Does MP, van den Elzen PJM, Hooykaas PJJ. Extrachromosomal homologous recombination and gene targeting in plant cells after Agrobacterium mediated transformation. EMBO J , 1990, 9(10): 3077-3084.
[19] Halfter U, Morris PC, Willmitzer L. Gene targeting in Arabidopsis thaliana . Mol Gen Genet , 1992, 231(2): 186-193.
[20] Swoboda P, Gal S, Hohn B, Puchta H. Intrachromosomal homologous recombination in whole plants. EMBO J , 1994, 13(2): 484-489.
[21] Miao ZH, Lam E. Targeted disruption of the TGA3 locus in Arabidopsis thaliana . Plant J , 1995, 7(2): 359-365.
[22] Puchta H, Dujon B, Hohn B. Homologous recombination in plant cells is enhanced by in vivo induction of double strand breaks into DNA by a site-specific endonuclease. Nucleic Acids Res , 1993, 21(22): 5034-5040.
[23] Puchta H, Dujon B, Hohn B. Two different but related mechanisms are used in plants for the repair of genomic double-strand breaks by homologous recombination. Proc Natl Acad Sci USA , 1996, 93(10): 5055-5060.
[24] Fauser F, Roth N, Pacher M, Ilg G, Sánchez-Fernández R, Biesgen C, Puchta H. In planta gene targeting. Proc Natl Acad Sci USA , 2012, 109(19): 7535-7540.
[25] Wright DA, Townsend JA, Winfrey RJ, Jr., Irwin PA, Rajagopal J, Lonosky PM, Hall BD, Jondle MD, Voytas DF. High-frequency homologo
