CRISPR-Barcoding for Intratumor Genetic Heterogeneity Modeling and Functional Analysis of Oncogenic Driver Mutations.
CRISPR-Barcoding for Intratumor Genetic Heterogeneity Modeling and Functional Analysis of Oncogenic Driver Mutations.
复制标题
DOI：
10.1016/j.molcel.2016.06.017
复制
发表时间：
2016-08-04
期刊：
影响因子：
16
通讯作者：
Grumolato L
中科院分区：
文献类型：
--
作者：
Guernet A;Mungamuri SK;Cartier D;Sachidanandam R;Jayaprakash A;Adriouch S;Vezain M;Charbonnier F;Rohkin G;Coutant S;Yao S;Ainani H;Alexandre D;Tournier I;Boyer O;Aaronson SA;Anouar Y;Grumolato L
肿瘤内部的遗传异质性是肿瘤进化和适应不同环境条件的基础。利用CRISPR/Cas9技术和特定的DNA条形码，我们设计了一种策略来概括和追踪含有感兴趣突变的癌细胞亚群的出现。我们使用这种方法来模拟肺癌细胞对EGFR抑制剂耐药的不同机制，并评估联合药物治疗的效果。通过克服当前方法的内在局限性，crispr条形码还可以研究大多数类型的遗传修饰，包括致癌驱动突变的修复。最后，我们使用高度复杂的条形码插入到特定的基因组位置，作为同时追踪数千个遗传标记癌细胞命运的手段。crispr条形码是一种直接且高度灵活的方法，在密切模仿癌症复杂性的背景下，它将极大地促进对特定突变的功能研究。Guernet等人利用CRISPR/Cas9技术，通过在特定基因组位置插入一系列点突变，对大量群体中的肿瘤细胞进行遗传标记和追踪。特定实验条件对条形码细胞片段的影响可以通过qPCR或深度测序来评估。
Intratumor genetic heterogeneity underlies the ability of tumors to evolve and adapt to different environmental conditions. Using CRISPR/Cas9 technology and specific DNA barcodes, we devised a strategy to recapitulate and trace the emergence of subpopulations of cancer cells containing a mutation of interest. We used this approach to model different mechanisms of lung cancer cell resistance to EGFR inhibitors and to assess effects of combined drug therapies. By overcoming intrinsic limitations of current approaches, CRISPR-barcoding also enables investigation of most types of genetic modifications, including repair of oncogenic driver mutations. Finally, we used highly complex barcodes inserted at a specific genome location as a means of simultaneously tracing the fates of many thousands of genetically labeled cancer cells. CRISPR-barcoding is a straightforward and highly flexible method that should greatly facilitate the functional investigation of specific mutations, in a context that closely mimics the complexity of cancer. Guernet et al. used CRISPR/Cas9 technology to genetically label and trace tumor cells within a mass population through insertion of a series of point mutations at a specific genomic location. The effects of a particular experimental condition on the fraction of barcoded cells can be assessed by qPCR or deep sequencing.
登录
查看更多内容
影响因子：
64.8
作者：
Greaves, Mel;Maley, Carlo C.
通讯作者：
Maley, Carlo C.
影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F
影响因子：
158.5
作者：
Jaenne, Pasi A.;Yang, James Chih-Hsin;Ranson, Malcolm
通讯作者：
Ranson, Malcolm
影响因子：
64.8
作者：
Maddalo, Danilo;Manchado, Eusebio;Concepcion, Carla P.;Bonetti, Ciro;Vidigal, Joana A.;Han, Yoon-Chi;Ogrodowski, Paul;Crippa, Alessandra;Rekhtman, Natasha;de Stanchina, Elisa;Lowe, Scott W.;Ventura, Andrea
通讯作者：
Ventura, Andrea
影响因子：
82.9
作者：
Hata AN;Niederst MJ;Archibald HL;Gomez-Caraballo M;Siddiqui FM;Mulvey HE;Maruvka YE;Ji F;Bhang HE;Krishnamurthy Radhakrishna V;Siravegna G;Hu H;Raoof S;Lockerman E;Kalsy A;Lee D;Keating CL;Ruddy DA;Damon LJ;Crystal AS;Costa C;Piotrowska Z;Bardelli A;Iafrate AJ;Sadreyev RI;Stegmeier F;Getz G;Sequist LV;Faber AC;Engelman JA
通讯作者：
Engelman JA