秀丽隐杆线虫中CRISPR-Cas9引导的基因组工程
CRISPR-Cas9-Guided Genome Engineering in C. elegans.
作者信息
Kim Hyun-Min, Colaiácovo Monica P
机构信息
Department of Genetics, Harvard Medical School, Boston, Massachusetts.
出版信息
Curr Protoc Mol Biol. 2016 Jul 1;115:31.7.1-31.7.18. doi: 10.1002/cpmb.7.
Abstract
The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) system is successfully being used for efficient and targeted genome editing in various organisms, including the nematode C. elegans. Recent studies have developed various CRISPR-Cas9 approaches to enhance genome engineering via two major DNA double-strand break repair pathways: non-homologous end joining and homologous recombination. Here we describe a protocol for Cas9-mediated C. elegans genome editing together with single guide RNA (sgRNA) and repair template cloning, as well as injection methods required for delivering Cas9, sgRNAs, and repair template DNA into the C. elegans germline. © 2016 by John Wiley & Sons, Inc.
摘要
CRISPR（规律成簇间隔短回文重复序列）-Cas（CRISPR相关）系统已成功用于多种生物体的高效靶向基因组编辑，包括线虫秀丽隐杆线虫。最近的研究开发了各种CRISPR-Cas9方法，通过两种主要的DNA双链断裂修复途径来增强基因组工程：非同源末端连接和同源重组。本文我们描述了一种用于Cas9介导的秀丽隐杆线虫基因组编辑的方案，以及单向导RNA（sgRNA）和修复模板克隆，还有将Cas9、sgRNAs和修复模板DNA导入秀丽隐杆线虫种系所需的注射方法。© 2016约翰威立父子公司版权所有
相似文献
1
CRISPR-Cas9-Guided Genome Engineering in C. elegans.
Curr Protoc Mol Biol. 2016 Jul 1;115:31.7.1-31.7.18. doi: 10.1002/cpmb.7.
2
CRISPR-Cas9-Guided Genome Engineering in Caenorhabditis elegans.
Curr Protoc Mol Biol. 2019 Dec;129(1):e106. doi: 10.1002/cpmb.106.
3
Precision genome editing using CRISPR-Cas9 and linear repair templates in C. elegans.
Methods. 2017 May 15;121-122:86-93. doi: 10.1016/j.ymeth.2017.03.023. Epub 2017 Apr 7.
4
[CRISPR/CAS9, the King of Genome Editing Tools].
Mol Biol (Mosk). 2017 Jul-Aug;51(4):582-594. doi: 10.7868/S0026898417040036.
5
Design of a generic CRISPR-Cas9 approach using the same sgRNA to perform gene editing at distinct loci.
BMC Biotechnol. 2019 Mar 20;19(1):18. doi: 10.1186/s12896-019-0509-7.
6
Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination.
Nucleic Acids Res. 2013 Nov;41(20):e193. doi: 10.1093/nar/gkt805. Epub 2013 Sep 5.
7
[CRISPR-Cas9 mediated genome editing in Caenorhabditis elegans].
Sheng Wu Gong Cheng Xue Bao. 2017 Oct 25;33(10):1693-1699. doi: 10.13345/j.cjb.170177.
8
Using Microinjection to Generate Genetically Modified Caenorhabditis elegans by CRISPR/Cas9 Editing.
Methods Mol Biol. 2019;1874:431-457. doi: 10.1007/978-1-4939-8831-0_25.
9
Creating Genome Modifications in C. elegans Using the CRISPR/Cas9 System.
Methods Mol Biol. 2015;1327:59-74. doi: 10.1007/978-1-4939-2842-2_6.
10
CRISPR-Cas systems: ushering in the new genome editing era.
Bioengineered. 2018;9(1):214-221. doi: 10.1080/21655979.2018.1470720.
引用本文的文献
1
Conferring High IAA Productivity on Low-IAA-Producing Organisms with PonAAS2, an Aromatic Aldehyde Synthase of a Galling Sawfly, and Identification of Its Inhibitor.
Insects. 2023 Jul 2;14(7):598. doi: 10.3390/insects14070598.
2
GRAS-1 is a novel regulator of early meiotic chromosome dynamics in C. elegans.
PLoS Genet. 2023 Feb 21;19(2):e1010666. doi: 10.1371/journal.pgen.1010666. eCollection 2023 Feb.
3
Peel-1 negative selection promotes screening-free CRISPR-Cas9 genome editing in Caenorhabditis elegans.
PLoS One. 2020 Sep 22;15(9):e0238950. doi: 10.1371/journal.pone.0238950. eCollection 2020.
4
A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins.
J Vis Exp. 2018 Apr 30(134):57518. doi: 10.3791/57518.
5
Fanconi Anemia FANCM/FNCM-1 and FANCD2/FCD-2 Are Required for Maintaining Histone Methylation Levels and Interact with the Histone Demethylase LSD1/SPR-5 in .
Genetics. 2018 Jun;209(2):409-423. doi: 10.1534/genetics.118.300823. Epub 2018 Mar 27.
6
What Can We Learn About Human Disease from the Nematode C. elegans?
Methods Mol Biol. 2018;1706:53-75. doi: 10.1007/978-1-4939-7471-9_4.
7
Precision genome editing using CRISPR-Cas9 and linear repair templates in C. elegans.
Methods. 2017 May 15;121-122:86-93. doi: 10.1016/j.ymeth.2017.03.023. Epub 2017 Apr 7.
本文引用的文献
1
DNA Damage Sensitivity Assays in .
Bio Protoc. 2015;5(11). Epub 2015 Jun 5.
2
Efficient Genome Editing in Caenorhabditis elegans with a Toolkit of Dual-Marker Selection Cassettes.
Genetics. 2015 Oct;201(2):449-58. doi: 10.1534/genetics.115.180679. Epub 2015 Jul 30.
3
Streamlined Genome Engineering with a Self-Excising Drug Selection Cassette.
Genetics. 2015 Aug;200(4):1035-49. doi: 10.1534/genetics.115.178335. Epub 2015 Jun 3.
4
New Insights into the Post-Translational Regulation of DNA Damage Response and Double-Strand Break Repair in Caenorhabditis elegans.
Genetics. 2015 Jun;200(2):495-504. doi: 10.1534/genetics.115.175661. Epub 2015 Mar 26.
5
Dramatic enhancement of genome editing by CRISPR/Cas9 through improved guide RNA design.
Genetics. 2015 Apr;199(4):959-71. doi: 10.1534/genetics.115.175166. Epub 2015 Feb 18.
6
Rapid and precise engineering of the Caenorhabditis elegans genome with lethal mutation co-conversion and inactivation of NHEJ repair.
Genetics. 2015 Feb;199(2):363-77. doi: 10.1534/genetics.114.172361. Epub 2014 Dec 9.
7
Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans.
Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23.
8
Efficient marker-free recovery of custom genetic modifications with CRISPR/Cas9 in Caenorhabditis elegans.
Genetics. 2014 Nov;198(3):837-46. doi: 10.1534/genetics.114.169730. Epub 2014 Aug 26.
9
A co-CRISPR strategy for efficient genome editing in Caenorhabditis elegans.
Genetics. 2014 Aug;197(4):1069-80. doi: 10.1534/genetics.114.166389. Epub 2014 May 30.
10
Engineering the Caenorhabditis elegans genome using Cas9-triggered homologous recombination.
Nat Methods. 2013 Oct;10(10):1028-34. doi: 10.1038/nmeth.2641. Epub 2013 Sep 1.
Zotero 插件