用于基因组编辑和癌症建模的CRISPR-Cas9基因敲入小鼠
CRISPR-Cas9 knockin mice for genome editing and cancer modeling.
作者信息
Platt Randall J, Chen Sidi, Zhou Yang, Yim Michael J, Swiech Lukasz, Kempton Hannah R, Dahlman James E, Parnas Oren, Eisenhaure Thomas M, Jovanovic Marko, Graham Daniel B, Jhunjhunwala Siddharth, Heidenreich Matthias, Xavier Ramnik J, Langer Robert, Anderson Daniel G, Hacohen Nir, Regev Aviv, Feng Guoping, Sharp Phillip A, Zhang Feng
机构信息
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
出版信息
Cell. 2014 Oct 9;159(2):440-55. doi: 10.1016/j.cell.2014.09.014. Epub 2014 Sep 25.
CRISPR-Cas9 is a versatile genome editing technology for studying the functions of genetic elements. To broadly enable the application of Cas9 in vivo, we established a Cre-dependent Cas9 knockin mouse. We demonstrated in vivo as well as ex vivo genome editing using adeno-associated virus (AAV)-, lentivirus-, or particle-mediated delivery of guide RNA in neurons, immune cells, and endothelial cells. Using these mice, we simultaneously modeled the dynamics of KRAS, p53, and LKB1, the top three significantly mutated genes in lung adenocarcinoma. Delivery of a single AAV vector in the lung generated loss-of-function mutations in p53 and Lkb1, as well as homology-directed repair-mediated Kras(G12D) mutations, leading to macroscopic tumors of adenocarcinoma pathology. Together, these results suggest that Cas9 mice empower a wide range of biological and disease modeling applications.
CRISPR-Cas9是一种用于研究遗传元件功能的多功能基因组编辑技术。为了广泛实现Cas9在体内的应用，我们建立了一种依赖Cre的Cas9基因敲入小鼠。我们使用腺相关病毒（AAV）、慢病毒或颗粒介导的引导RNA在神经元、免疫细胞和内皮细胞中进行了体内和体外基因组编辑的演示。利用这些小鼠，我们同时模拟了肺腺癌中三个显著突变的基因KRAS、p53和LKB1的动态变化。在肺中递送单个AAV载体可产生p53和Lkb1的功能丧失突变，以及同源定向修复介导的Kras（G12D）突变，从而导致腺癌病理的宏观肿瘤。总之，这些结果表明Cas9小鼠可用于广泛的生物学和疾病建模应用。
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