Cell子刊:CRISPR筛查鉴定大肠癌耐药驱动因子
创作:爱的抉择 审核:Lexi 01月06日
  • 新方法通过捕获单个类器官中sgRNA的整合,提高CRISPR筛选的准确性和稳定性,减少基因组规模筛选需要的细胞数量;
  • TGF-β介导的生长限制是大肠癌进展的一个关键过程;
  • 对野生型和APC突变型人肠道类器官进行CRISPR筛选,鉴定出与TGF-β耐药性相关基因,其中包括肿瘤抑制性SWI/SNF染色质重塑复合物的多个亚单位;
  • 这些基因突变需要APC的持续失活来促进TGF-β耐药性和削弱TGF-β靶基因的转录;
  • SWI/SNF复合物控制TGF-β靶基因的可接近性。
主编推荐语
爱的抉择
利用CRISPR进行全基因组的正向遗传筛选是研究细胞环路和信号通路的有力工具,但将其应用于类器官的研究却受到技术限制的阻碍。发表在Cell Stem Cell的研究开发了一种利用全基因组CRISPR在人类肠道类器官中筛选肿瘤信号通路的方法。一些抑癌基因被确定为APC突变背景下TGF-β通路的负调控因子,包括SWI/SNF染色质重塑复合体的亚基。
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Cell Stem Cell [IF:20.86]

Genome-Scale CRISPR Screening in Human Intestinal Organoids Identifies Drivers of TGF-β Resistance

在人类肠道类器官中进行基因组CRISPR筛查以鉴定TGF-β耐药性的驱动因子

10.1016/j.stem.2020.02.007

2020-03-05, Other

Abstract & Authors:展开

Abstract:收起
Forward genetic screens with genome-wide CRISPR libraries are powerful tools for resolving cellular circuits and signaling pathways. Applying this technology to organoids, however, has been hampered by technical limitations. Here we report improved accuracy and robustness for pooled-library CRISPR screens by capturing sgRNA integrations in single organoids, substantially reducing required cell numbers for genome-scale screening. We applied our approach to wild-type and APC mutant human intestinal organoids to identify genes involved in resistance to TGF-β-mediated growth restriction, a key process during colorectal cancer progression, and validated hits including multiple subunits of the tumor-suppressive SWI/SNF chromatin remodeling complex. Mutations within these genes require concurrent inactivation of APC to promote TGF-β resistance and attenuate TGF-β target gene transcription. Our approach can be applied to a variety of assays and organoid types to facilitate biological discovery in primary 3D tissue models.

First Authors:
Till Ringel

Correspondence Authors:
Gerald Schwank

All Authors:
Till Ringel,Nina Frey,Femke Ringnalda,Sharan Janjuha,Sarah Cherkaoui,Stefan Butz,Sumana Srivatsa,Martin Pirkl,Giancarlo Russo,Lukas Villiger,Gerhard Rogler,Hans Clevers,Niko Beerenwinkel,Nicola Zamboni,Tuncay Baubec,Gerald Schwank

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