德国癌症研究中心(DKFZ)Henner F. Farin研究组利用体内外CRISPR-Cas9汇集筛选,鉴定出人结肠类器官中的肿瘤抑制因子。2020年4月28日,《细胞—干细胞》在线发表了这一成果。
Title: Pooled In Vitro and In Vivo CRISPR-Cas9 Screening Identifies Tumor Suppressors in Human Colon Organoids
Author: Birgitta E. Michels, Mohammed H. Mosa, Barbara I. Streibl, Tianzuo Zhan, Constantin Menche, Khalil Abou-El-Ardat, Tahmineh Darvishi, Ewelina Czonka, Sebastian Wagner, Jan Winter, Hind Medyouf, Michael Boutros, Henner F. Farin
Issue&Volume: 2020-04-28
Abstract: Colorectal cancer (CRC) is characterized by prominent genetic and phenotypic heterogeneitybetween patients. To facilitate high-throughput genetic testing and functional identificationof tumor drivers, we developed a platform for pooled CRISPR-Cas9 screening in humancolon organoids. Using transforming growth factor β (TGF-β) resistance as a paradigmto establish sensitivity and scalability in vitro, we identified optimal conditions and strict guide RNA (gRNA) requirements for screeningin 3D organoids. We then screened a pan-cancer tumor suppressor gene (TSG) libraryin pre-malignant organoids with APC/;KRASG12D mutations, which were xenografted to study clonal advantages in context of a complextumor microenvironment. We identified TGFBR2 as the most prevalent TSG, followed by known and previously uncharacterized mediatorsof CRC growth. gRNAs were validated in a secondary screen using unique molecular identifiers(UMIs) to adjust for clonal drift and to distinguish clone size and abundance. Together,these findings highlight a powerful organoid-based platform for pooled CRISPR-Cas9screening for patient-specific functional genomics.
DOI: 10.1016/j.stem.2020.04.003
Source: https://www.cell.com/cell-stem-cell/fulltext/S1934-5909(20)30142-9
Cell Stem Cell:《细胞—干细胞》,创刊于2007年。隶属于细胞出版社,最新IF:21.464
官方网址:https://www.cell.com/cell-stem-cell/home
投稿链接:https://www.editorialmanager.com/cell-stem-cell/default.aspx