中国农业科学Issue(9)：1669-1677,9.DOI:10.3864/j.issn.0578-1752.2015.09.01
CRISPR/Cas9系统在植物基因组定点编辑中的研究进展
Progress in Research of CRISPR/Cas9 System in Genome Targeted Editing in Plants
摘要
Abstract
When exogenous DNA was imported into plant cell by transgenic technology, DNA fragment will integrate into the genome by homologous recombination or nonhomologous recombination. In addition, the plants seedling will achieve corresponding target traits. Homologous recombination occurred when the exogenous DNA and the same or similar sequences in receptor cells recombined and integrated to the receptor cell’s chromosomes, so the sequence will be possible to precisely modified and transformed. However, in some cases, to avoid the fracture caused by DNA or chromosomal DNA degradation or the impact on the vitality, the two double-stranded DNA break ends will be joined without considering the sequence similarity by error-prone nonhomologous end joining. As a result, precise mutations control is more difficult to achieve because insertion and/or deletion and other variety of mutations are high-frequently occurred in non-homologous recombination than in homologous recombination. Unfortunately, the frequency of homologous recombination is very low in plants which results the undesirable genome editing. Site-specific nucleases make genome editing more efficient and more precise by the great improvement of the efficiency in the homologous recombination. Such nucleases, zinc finger nuclease (ZFN) and transcription activator-like effector nucleases (TALENs), have been demonstrated to efficiently produce a DNA double-strand break at target site and to induce genome modification in a variety of organisms including plants. However, some defects found in the specific application, such as off-target effects, specific combined site with genome affected by chromosomal location and adjacent sequence, furthermore, technical complexity and time-consuming for assemble limit its application. The clustered, regularly interspaced, short palindromic repeats(CRISPR) system is a prokaryotic adaptive immune system which widely exists in bacteria and archaea. As the result of the long-term evolution, the system can defense against the degradation of RNA to guide the invasion of the virus or phage DNA. Recent advances in the study demonstrated CRISPR/Cas typeⅡ system was a promising system of genome editing strategy with high efficiency, affordability and easy to engineer compared with ZFNs and TALENs. Many precise genome editing cases by CRISPR/Cas system were found successfully in human cells, zebra fish, mice and bacterial, including gene insertion, deletion, mutation in multiple sites or in specific locus. Although the application in plants is still in a relatively limited range, there is an outstanding prospect of CRISPR/Cas system in plant genetic engineering. A brief summary of the composition and principles was presented firstly. Then, the authors emphasis on citing numerous cases involved its application of exogenous and endogenous genes editing in cotyledon and dicotyledonous plants which demonstrate that the CRISPR/Cas9 system has become a powerful tool in plant genome engineering. Finally, the future of the genome technology application in agriculture and plant genetic engineering was discussed which will provide a reference for researchers in genetic modification.关键词
CRISPR/Cas系统/基因组编辑/植物基因工程Key words
CRISPR/Cas system/genome editing/plant genetic engineering引用本文复制引用
解莉楠,宋凤艳,张旸..CRISPR/Cas9系统在植物基因组定点编辑中的研究进展[J].中国农业科学,2015,(9):1669-1677,9.基金项目
黑龙江省自然科学基金（C201343）、中央高校基本科研业务费专项资金（2572014CA21）、哈尔滨市科技创新人才研究专项资金（2013RFQXJ036）、黑龙江省博士后资助（LBH-Z14009）、“十二五”863计划现代农业技术领域项目 （）
