CRISPR/Cas系统在基因修饰植物及其产品检测应用中的原理和进展
第一作者：wangweixia@caas.cn
收稿日期: 2023-06-17
网络出版日期: 2023-11-21
基金资助
中央级公益性科研院所基本科研业务费专项(CPSIBRF-CNRRI-202122)
Principle and Progress of CRISPR/Cas System in the Detection of Genetically Modified Plants and Their Products
1st author: wangweixia@caas.cn
Received date: 2023-06-17
Online published: 2023-11-21
转基因及基因编辑的基因修饰植物发展迅猛，相关的检测技术也面临更多挑战，尤其是对于无外源DNA引入的基因编辑产品，其检测难度更高，传统的检测技术已无法满足。基于CRISPR/Cas检测系统中不同Cas蛋白所具有的附属非特异切割活性被广泛应用于分子检测领域，它与LAMP和RPA等温扩增相结合可实现准确快速的现场检测，甚至是对单核苷酸突变的基因编辑产品识别和检测。但是，该检测系统在植物基因修饰产品检测中的应用才刚刚起步，本文对该检测系统的原理及其在基因修饰的植物及其产品中的应用进行了评述。
关键词： CRISPR/Cas系统; 分子检测; 转基因; 基因编辑; 基因修饰植物
王渭霞, 朱廷恒, 赖凤香, 万品俊, 魏琪, 傅强 . CRISPR/Cas系统在基因修饰植物及其产品检测应用中的原理和进展[J]. 中国稻米, 2023 , 29(6) : 21 -27 . DOI: 10.3969/j.issn.1006-8082.2023.06.005
With the rapid development of genetically modified plants including transgenic and gene-edited plants, the related detection technologies are also facing more challenges, especially for gene editing products without the introduction of exogenous DNA, its detection is more difficult, and the traditional detection technology cannot be satisfied. The accessory nonspecific cleavage activity of different Cas proteins in the CRISPR-Cas-based detection system is widely used in the field of molecular detection. It can be combined with LAMP and RPA isothermal amplification to achieve accurate and rapid on-site detection, even for single gene editing products for nucleotide mutations. However, the application of the detection system in the detection of plant genetically modified products has just started. This paper reviews the principle of the detection system and its application in genetically modified plants and their products.
| [1] | BOLOTIN A, QUINQUIS B, SOROKIN A, et al. Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin[J]. Microbiology, 2005, 151 (8): 2 551-2 561. |
| [2] | MARRAFFIN L A. CRISPR-Cas-mediated base editing: Technical considerations and practical applications[J]. Trend Biotechnology, 2019, 37(10): 1 121-1 142. |
| [3] | GARNEAU J E, DUPUIS M E, VILLION M, et al. The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA[J]. Nature, 2010, 468(7320): 67-71. |
| [4] | HORVATH P, BARRANGOU R. Crispr/cas, the immune system of bacteria and archaea[J]. Science, 2010, 327(5962): 167-170. |
| [5] | ZHU H C, LI C, GAO C X. Applications of CRISPR-Cas in agriculture and plant biotechnology[J]. Nature Reviews Molecular Cell Biology, 2020, 21: 661-677. |
| [6] | CHEN K, GAO C. Targeted genome modification technologies and their applications in crop improvements[J]. Plant Cell Reports, 2014, 33(4): 575-583. |
| [7] | CHERTOW D S. Next-generation diagnostics with crispr[J]. Science, 2018, 360(6387): 381-382. |
| [8] | KHAMBHATI K, BHATTACHARJEE G, SINGH V. Current progress in crispr-based diagnostic platforms[J]. Journal of Cell Biochemistry, 2019, 120(3): 2 721-2 725. |
| [9] | WANG X, SHANG X Y, HUANG X X. Next-generation pathogen diagnosis with CRISPR/Cas-based detection methods[J]. Emerging Microbes & Infections, 2020, 9(1): 1 682-1 691. |
| [10] | ISHINO Y, SHINAGAWA H, MAKINO K, et al. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product[J]. Journal of Bacteriology, 1987, 169(12): 5 429-5 433. |
| [11] | JANSEN R, EMBDEN J D, GAASTRA W, et al. Identification of genes that are associated with DNA repeats in prokaryotes[J]. Molecular Microbiology, 2002, 43(6): 1 565-1 575. |
| [12] | BARRANGOU R, FREMAUX C, DEVEAU H, et al. CRISPR provides acquired resistance against viruses in prokaryotes[J]. Science, 2007, 315(5819): 1 709-1 712. |
| [13] | MARRAFFINI L A, SONTHEIMER E J. CRISPR interference limits horizontal gene transfer in staphylococci by targeting DNA[J]. Science, 2008, 322(5909): 1 843-1 845. |
| [14] | 赵海卫, 吕欣. CRISPR/Cas9系统靶向基因组编辑的新策略[J]. 中国病原微生物杂志, 2015, 10(3):281-284. |
| [15] | MAKAROVA K S, WOLF Y I, ALKHNBASHI O S, et al. An updated evolutionary classification of CRISPR-Cas systems[J]. Nature Review of Microbiology, 2015, 13(11): 722-736. |
| [16] | JIANG W Z, ZHOU H B, BI H H, et al. Demonstration of CRISPR/Cas9 sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice[J]. Nucleic Acids Research, 2013. |
| [17] | MAKAROVA K S, HAFT D H, BARRANGOU R, et al. Evolution and classification of the CRISPR-Cas systems[J]. Nature Review of Microbiology, 2011, 9 (6): 467-477. |
| [18] | KOMOR A C, KIM Y B, PACKER M S, et al. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage[J]. Nature, 2016, 533(7603): 420-424. |
| [19] | SHIMATANI Z, KASHOJIYA S, TAKAYAMA M, et al. Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion[J]. Nature Biotechnology, 2017, 35: 441-443. |
| [20] | GOOTENBERG J S, ABUDAYYEH O O, LEE J W, et al. Nucleic acid detection with CRISPR-Cas13a/c2c2[J]. Science, 2017, 356(6336): 438-442. |
| [21] | CHEN J S, MA E, HARRINGTON L B, et al. Crispr-Cas12a target binding unleashes indiscriminate single-stranded Dnase activity. Science, 2018, 360(6387): 436-439. |
| [22] | HARRINGTON L B, BURSTEIN D, CHEN J S, et al. Programmed DNA destruction by miniature CRISPR-Cas14 enzymes[J]. Science, 2018, 362(6416): 839-842. |
| [23] | LI S Y, CHENG Q X, WANG J M, et al. Crispr-cas12a-assisted nucleic acid detection[J]. Cell Discovery, 2018. |
| [24] | MOTA D S, MARQUES J M, GUIMARAES J M, et al. Research article CRISPR/Cas class 2 systems and their applications in biotechnological processes[J]. Genetics and Molecular Research, 2020, 19(1): 1-11. |
| [25] | LI Y, Li S W, WANG J, et al. Crispr/cas systems towards next-generation biosensing[J]. Trends Biotechnology, 2019, 37(7): 730-743. |
| [26] | EAST-SELETSKY A, O’CONNELL M R, KNIGHT S C, et al. Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection[J]. Nature, 2016, 538(7624): 270-273. |
| [27] | MYHRVOLD C, FREIJE C A, GOOTENBERG J S, et al. Field-deployable viral diagnostics using crispr-cas13[J]. Science, 2018, 360(6387): 444-448. |
| [28] | GOOTENBERG J S, ABUDAYYEH O O, KELLNER M J, et al. Multiplexed and portable nucleic acid detection platform with cas13, cas12a, and csm6[J]. Science, 2018, 360(6387): 439-444. |
| [29] | LI L, LI S, WU N, et al. HOLMESv2: a crispr-cas12b-assisted platform for nucleic acid detection and DNA methylation quantitation[J]. ACS Synthetic Biology, 2019, 8(10): 2 228-2 237. |
| [30] | AQUINO-JARQUIN G. CRISPR-Cas14 is now part of the artillery for gene editing and molecular diagnostic[J]. Nanomedicine : Nanotechnology, Biology, and Medicine, 2019, 18: 428-431. |
| [31] | NOTOMI T, OKAYAMA H, MASUBUCHI H, et al. Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Research, 2000, 28(12): e63. |
| [32] | DAHER R K, STEWART G, BOISSINOT M, et al. Recombinase polymerase amplification for diagnostic applications[J]. Clinical Chemistry, 2016, 62(7): 947-958. |
| [33] | ZHANG Y M, ZHANG Y, XIE K B. Evaluation of CRISPR/Cas12a-based DNA detection for fast pathogen diagnosis and GMO test in rice[J]. Molecular Breeding, 2020. |
| [34] | WU H, HE J S, ZHANG F, et al. Contamination-free visual detection of CaMV35S promoter amplicon using CRISPR/Cas12a coupled with a designed reaction vessel: Rapid, specific and sensitive[J]. Analytica Chimica Acta, 2020, 1096: 130-137. |
| [35] | 潘志文, 张旭冬, 高洁儿, 等. 基因组编辑植物的监管与检测技术[J]. 科技导报, 2021, 39(9):87-92. |
| [36] | LIU Q, JIAO X Z, MENG X B, et al. FED: A web tool for foreign element detection of genome-edited organism[J]. Science China Life Sciences, 2020, 63(1): 167-170. |
| [37] | 王梦雨, 王灏潜, 王旭静, 等. 基因编辑产品检测技术研究进展[J]. 生物技术进展, 2021, 11(4):438-445. |
| [38] | PENG C, ZHENG M, DING L, et al. Accurate detection and evaluation of the gene-editing frequency in plants using droplet digital PCR[J]. Frontiers in Plant Science, 2021. |
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