使用CRISPR - Cas9介导的基因组编辑生成小鼠模型

Q1 Agricultural and Biological Sciences

Current protocols in mouse biology Pub Date : 2016-03-01 DOI:10.1002/9780470942390.mo150178

Wenning Qin, Peter M. Kutny, R. Maser, S. Dion, Jeffrey D. Lamont, Yingfan Zhang, Greggory A. Perry, Haoyi Wang

{"title":"使用CRISPR - Cas9介导的基因组编辑生成小鼠模型","authors":"Wenning Qin, Peter M. Kutny, R. Maser, S. Dion, Jeffrey D. Lamont, Yingfan Zhang, Greggory A. Perry, Haoyi Wang","doi":"10.1002/9780470942390.mo150178","DOIUrl":null,"url":null,"abstract":"The CRISPR‐Cas9 system in bacteria and archaea has recently been exploited for genome editing in various model organisms, including mice. The CRISPR‐Cas9 reagents can be delivered directly into the mouse zygote to derive a mutant animal carrying targeted genetic modifications. The major components of the system include the guide RNA, which provides target specificity, the Cas9 nuclease that creates the DNA double‐strand break, and the donor oligonucleotide or plasmid carrying the intended mutation flanked by sequences homologous to the target site. Here we describe the general considerations and experimental protocols for creating genetically modified mice using the CRISPR‐Cas9 system. © 2016 by John Wiley & Sons, Inc.","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"28 2","pages":"39 - 66"},"PeriodicalIF":0.0000,"publicationDate":"2016-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/9780470942390.mo150178","citationCount":"42","resultStr":"{\"title\":\"Generating Mouse Models Using CRISPR‐Cas9‐Mediated Genome Editing\",\"authors\":\"Wenning Qin, Peter M. Kutny, R. Maser, S. Dion, Jeffrey D. Lamont, Yingfan Zhang, Greggory A. Perry, Haoyi Wang\",\"doi\":\"10.1002/9780470942390.mo150178\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The CRISPR‐Cas9 system in bacteria and archaea has recently been exploited for genome editing in various model organisms, including mice. The CRISPR‐Cas9 reagents can be delivered directly into the mouse zygote to derive a mutant animal carrying targeted genetic modifications. The major components of the system include the guide RNA, which provides target specificity, the Cas9 nuclease that creates the DNA double‐strand break, and the donor oligonucleotide or plasmid carrying the intended mutation flanked by sequences homologous to the target site. Here we describe the general considerations and experimental protocols for creating genetically modified mice using the CRISPR‐Cas9 system. © 2016 by John Wiley & Sons, Inc.\",\"PeriodicalId\":37980,\"journal\":{\"name\":\"Current protocols in mouse biology\",\"volume\":\"28 2\",\"pages\":\"39 - 66\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/9780470942390.mo150178\",\"citationCount\":\"42\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current protocols in mouse biology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1002/9780470942390.mo150178\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"Agricultural and Biological Sciences\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in mouse biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1002/9780470942390.mo150178","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Agricultural and Biological Sciences","Score":null,"Total":0}

引用次数: 42

摘要

细菌和古细菌中的CRISPR‐Cas9系统最近被用于多种模式生物(包括小鼠)的基因组编辑。CRISPR‐Cas9试剂可以直接传递到小鼠受精卵中，以获得携带靶向基因修饰的突变动物。该系统的主要组成部分包括提供目标特异性的引导RNA，产生DNA双链断裂的Cas9核酸酶，以及携带预期突变的供体寡核苷酸或质粒，其两侧是与目标位点同源的序列。在这里，我们描述了使用CRISPR‐Cas9系统创建转基因小鼠的一般考虑因素和实验方案。©2016 by John Wiley & Sons, Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

Generating Mouse Models Using CRISPR‐Cas9‐Mediated Genome Editing

The CRISPR‐Cas9 system in bacteria and archaea has recently been exploited for genome editing in various model organisms, including mice. The CRISPR‐Cas9 reagents can be delivered directly into the mouse zygote to derive a mutant animal carrying targeted genetic modifications. The major components of the system include the guide RNA, which provides target specificity, the Cas9 nuclease that creates the DNA double‐strand break, and the donor oligonucleotide or plasmid carrying the intended mutation flanked by sequences homologous to the target site. Here we describe the general considerations and experimental protocols for creating genetically modified mice using the CRISPR‐Cas9 system. © 2016 by John Wiley & Sons, Inc.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

自引率

0.00%

发文量

期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.