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A Sleeping Beauty transposon-mediated screen identifies murine susceptibility genes for adenomatous polyposis coli (Apc)-dependent intestinal tumorigenesis
Journal article   Open access   Peer reviewed

A Sleeping Beauty transposon-mediated screen identifies murine susceptibility genes for adenomatous polyposis coli (Apc)-dependent intestinal tumorigenesis

Timothy K Starr, Patricia M Scott, Benjamin M Marsh, Lei Zhao, Bich L. N Than, M. Gerard O'Sullivan, Aaron L Sarver, Adam J Dupuy, David A Largaespada and Robert T Cormier
Proceedings of the National Academy of Sciences - PNAS, Vol.108(14), pp.5765-5770
04/05/2011
DOI: 10.1073/pnas.1018012108
PMCID: PMC3078351
PMID: 21436051
url
https://doi.org/10.1073/pnas.1018012108View
Published (Version of record) Open Access

Abstract

It is proposed that a progressive series of mutations and epigenetic events leads to human colorectal cancer (CRC) and metastasis. Furthermore, data from resequencing of the coding regions of human CRC suggests that a relatively large number of mutations occur in individual human CRC, most at low frequency. The functional role of these low-frequency mutations in CRC, and specifically how they may cooperate with high-frequency mutations, is not well understood. One of the most common rate-limiting mutations in human CRC occurs in the adenomatous polyposis coli ( APC ) gene. To identify mutations that cooperate with mutant APC , we performed a forward genetic screen in mice carrying a mutant allele of Apc ( Apc Min ) using Sleeping Beauty ( SB ) transposon-mediated mutagenesis. Apc Min SB -mutagenized mice developed three times as many polyps as mice with the Apc Min allele alone. Analysis of transposon common insertion sites (CIS) identified the Apc locus as a major target of SB -induced mutagenesis, suggesting that SB insertions provide an efficient route to biallelic Apc inactivation. We also identified an additional 32 CIS genes/loci that may represent modifiers of the Apc Min phenotype. Five CIS genes tested for their role in proliferation caused a significant change in cell viability when message levels were reduced in human CRC cells. These findings demonstrate the utility of using transposon mutagenesis to identify low-frequency and cooperating cancer genes; this approach will aid in the development of combinatorial therapies targeting this deadly disease.
transgenic mice Biological Sciences cancer gene discovery

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