TY - JOUR
T1 - CRISPR-Cas9 knockin mice for genome editing and cancer modeling
AU - Platt, Randall J.
AU - Chen, Sidi
AU - Zhou, Yang
AU - Yim, Michael J.
AU - Swiech, Lukasz
AU - Kempton, Hannah R.
AU - Dahlman, James E.
AU - Parnas, Oren
AU - Eisenhaure, Thomas M.
AU - Jovanovic, Marko
AU - Graham, Daniel B.
AU - Jhunjhunwala, Siddharth
AU - Heidenreich, Matthias
AU - Xavier, Ramnik J.
AU - Langer, Robert
AU - Anderson, Daniel G.
AU - Hacohen, Nir
AU - Regev, Aviv
AU - Feng, Guoping
AU - Sharp, Phillip A.
AU - Zhang, Feng
N1 - Funding Information: We thank the entire Zhang laboratory and Sharp laboratory. We thank J.T. Ting, I. Tirosh, N. Sanjana, S. Jones, Y. Li, and colleagues in the Broad and Koch Institutes for technical assistance and discussions. We thank the Swanson Biotechnology Center for their support (Applied Therapeutics and Whole Animal Imaging, Bioinformatics and Computing, ES Cell and Transgenics, Microscopy, Flow Cytometry, and Histology, in particular). R.J.P. is supported by a National Science Foundation Graduate Research Fellowship under grant number 1122374. S.C. is a Damon Runyon Cancer Research Fellow (DRG-2117-12). Y.Z. is supported by the Simons Center for the Social Brain at MIT, postdoctoral fellowship. L.S. is a European Molecular Biology Organization (EMBO) and Foundation for Polish Science (FNP) Fellow. J.E.D. is supported by the NDSEG, NSF, and MIT Presidential Graduate Fellowships. M.J. is supported by fellowships of the Swiss National Science Foundation for advanced researchers (SNF) and the Marie Sklodowska-Curie IOF. R.J.X. and D.G. are supported by the Helmsley CharitableTrust and DK43351. M.H. is supported by a postdoctoral fellowship from the Human Frontiers Science Program. A.R. is supported by NHGRI CEGS P50 HG006193, HHMI and the Klarman Cell Observatory. N.H. is supported by NHGRI CEGS P50 HG006193. R.L. and D.A.G. are supported by the NIH Centers of Cancer Nanotechnology Excellence grant U54CA151884, the Controlled Release grant EB000244, and the National Heart, Lung, and Blood Institute, National Institutes of Health, as a Program of Excellence in Nanotechnology (PEN) Award, contract HHSN268201000045C. G.F. is supported by the McGovern Internal Funding Poitras Gift 1631119, the Stanley Center, the SFARI/Simons Foundation 6927482, and the Nancy Lurie Marks Family Foundation 6928117. P.A.S. is supported by United States Public Health Service R01-CA133404 from the National Institutes of Health; by an MIT-Harvard Center for Cancer Nanotechnology Excellence Grant U54 CA151884 from the National Cancer Institute; by a generous gift from the Marie D. and Pierre Casimir-Lambert Fund; by an SkTech/MIT Initiative Grant from the Skolkovo Foundation; and partially by the Koch Institute Support (core) grant P30-CA14051 from the National Cancer Institute. F.Z. is supported by the NIMH through a NIH Director’s Pioneer Award (DP1-MH100706), the NINDS through a NIH Transformative R01 grant (R01-NS 07312401), NSF Waterman Award, the Keck, Damon Runyon, Searle Scholars, Klingenstein, Vallee, Merkin, and Simons Foundations, and Bob Metcalfe. CRISPR reagents are available to the academic community through Addgene, and associated protocols, support forum, and computational tools are available via the Zhang lab website ( http://www.genome-engineering.org ). The Cre-dependent Cas9 (JAX Stock Number: 024857) and constitutive Cas9-expressing (JAX Stock Number: 024858) mice are available via the Jackson Laboratory. Publisher Copyright: © 2014 Elsevier Inc.
PY - 2014/10/9
Y1 - 2014/10/9
N2 - 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 KrasG12D 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.
AB - 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 KrasG12D 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.
UR - http://www.scopus.com/inward/record.url?scp=84912101598&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.cell.2014.09.014
DO - https://doi.org/10.1016/j.cell.2014.09.014
M3 - Article
C2 - 25263330
SN - 0092-8674
VL - 159
SP - 440
EP - 455
JO - Cell
JF - Cell
IS - 2
ER -
