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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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超级Cas12a能够实现高度复杂的表观基因组编辑屏幕.

Schuyler M Melore^1,2,3,4, Marisa C Hamilton^1,3,4, Timothy E Reddy^1,2,3,4

¹University Program in Genetics & Genomics, Duke University, Durham, NC, USA.

bioRxiv : the preprint server for biology

|July 19, 2024

概括

研究人员开发了一种新的基于CRISPR的表观基因组编辑工具,用于高度多重化屏幕. 这种系统能够同时激活和抑制基因,以研究复杂的遗传相互作用和基因调节.

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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

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Published on: December 11, 2020

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Published on: December 11, 2020

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
表观遗传学在表观遗传学中,表观遗传学是指表观遗传学.

背景情况:

基因和 cis 调节元素 (CRE) 相互作用对生物过程和疾病至关重要.
以前的表观基因组编辑工具仅限于对对基因相互作用分析.

研究的目的:

开发一种高度多重化的表观基因组编辑系统,用于评估复杂的遗传相互作用.
为了使综合基因调节研究能够同时激活和抑制基因.

主要方法:

利用一个高效的dCas12a (dHyperLbCas12a) 系统与长的CRISPRRNA (crRNA) 数组进行多重表观基因组编辑.
与各种激活 (P300) 和抑制 (SIN3A交互域 - SID) 域证明了兼容性.
在单个crRNA阵列上使用共同表达的dCas12a正义词启用了同时激活和抑制.

主要成果:

该dCas12a平台支持高度多重的表观基因组编辑和同时激活/抑制基因.
成功选了约19,000条条码crRNA数组,以分析CRE对基因表达的贡献.
剖析了CREs对剂量依赖基因表达控制的独立和组合效应.

结论:

开发的dCas12a系统提供了高效,高度多重化的表观基因组编辑功能.
这项技术扩大了研究基因调节和各种生物系统中的基因相互作用的可能性.
为高通量基因选提供了一个强大的新工具.