TY - JOUR
T1 - CRISPR/Cas9 somatic multiplex-mutagenesis for high-Throughput functional cancer genomics in mice
AU - Weber, Julia
AU - Öllinger, Rupert
AU - Friedrich, Mathias
AU - Ehmer, Ursula
AU - Barenboim, Maxim
AU - Steiger, Katja
AU - Heid, Irina
AU - Mueller, Sebastian
AU - Maresch, Roman
AU - Engleitner, Thomas
AU - Gross, Nina
AU - Geumann, Ulf
AU - Fu, Beiyuan
AU - Segler, Angela
AU - Yuan, Detian
AU - Lange, Sebastian
AU - Strong, Alexander
AU - Rosa, Jorge De La
AU - Esposito, Irene
AU - Liu, Pentao
AU - Cadiñanos, Juan
AU - Vassiliou, George S.
AU - Schmid, Roland M.
AU - Schneider, Gönter
AU - Unger, Kristian
AU - Yang, Fengtang
AU - Braren, Rickmer
AU - Heikenwälder, Mathias
AU - Varela, Ignacio
AU - Saur, Dieter
AU - Bradley, Allan
AU - Rad, Roland
PY - 2015/11/10
Y1 - 2015/11/10
N2 - Here, we show CRISPR/Cas9-based targeted somatic multiplexmutagenesis and its application for high-Throughput analysis of gene function in mice. Using hepatic single guide RNA (sgRNA) delivery, we targeted large gene sets to induce hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC). We observed Darwinian selection of target genes, which suppress tumorigenesis in the respective cellular/tissue context, such as Pten or Cdkn2a, and conversely found low frequency of Brca1/2 alterations, explaining mutational spectra in human ICC/HCC. Our studies show that multiplexed CRISPR/Cas9 can be used for recessive genetic screening or high-Throughput cancer gene validation in mice. The analysis of CRISPR/Cas9-induced tumors provided support for a major role of chromatin modifiers in hepatobiliary tumorigenesis, including that of ARID family proteins, which have recently been reported to be mutated in ICC/HCC. We have also comprehensively characterized the frequency and size of chromosomal alterations induced by combinatorial sgRNA delivery and describe related limitations of CRISPR/Cas9 multiplexing, as well as opportunities for chromosome engineering in the context of hepatobiliary tumorigenesis. Our study describes novel approaches to model and study cancer in a high-Throughput multiplexed format that will facilitate the functional annotation of cancer genomes.
AB - Here, we show CRISPR/Cas9-based targeted somatic multiplexmutagenesis and its application for high-Throughput analysis of gene function in mice. Using hepatic single guide RNA (sgRNA) delivery, we targeted large gene sets to induce hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC). We observed Darwinian selection of target genes, which suppress tumorigenesis in the respective cellular/tissue context, such as Pten or Cdkn2a, and conversely found low frequency of Brca1/2 alterations, explaining mutational spectra in human ICC/HCC. Our studies show that multiplexed CRISPR/Cas9 can be used for recessive genetic screening or high-Throughput cancer gene validation in mice. The analysis of CRISPR/Cas9-induced tumors provided support for a major role of chromatin modifiers in hepatobiliary tumorigenesis, including that of ARID family proteins, which have recently been reported to be mutated in ICC/HCC. We have also comprehensively characterized the frequency and size of chromosomal alterations induced by combinatorial sgRNA delivery and describe related limitations of CRISPR/Cas9 multiplexing, as well as opportunities for chromosome engineering in the context of hepatobiliary tumorigenesis. Our study describes novel approaches to model and study cancer in a high-Throughput multiplexed format that will facilitate the functional annotation of cancer genomes.
UR - http://www.scopus.com/inward/record.url?scp=84946771643&partnerID=8YFLogxK
U2 - 10.1073/pnas.1512392112
DO - 10.1073/pnas.1512392112
M3 - Article
C2 - 26508638
AN - SCOPUS:84946771643
SN - 0027-8424
VL - 112
SP - 13982
EP - 13987
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 45
ER -