Open AccessA Sleeping Beauty transposon-mediated screen identifies murine susceptibility genes for adenomatous polyposis coli ( Apc )-dependent intestinal tumorigenesis
Author(s) -
Timothy K. Starr,
Patricia M. Scott,
Benjamin M. Marsh,
Lei Zhao,
B L N Than,
M. Gerard O’Sullivan,
Aaron L. Sarver,
Adam J. Dupuy,
David A. Largaespada,
Robert T. Cormier
Publication year - 2011
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.1018012108
Subject(s) - adenomatous polyposis coli , biology , genetics , transposon mutagenesis , mutagenesis , sleeping beauty transposon system , gene , carcinogenesis , genetic screen , transposable element , mutant , mutation , allele , epigenetics , familial adenomatous polyposis , point mutation , colorectal cancer , cancer
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 mutantAPC , we performed a forward genetic screen in mice carrying a mutant allele ofApc (ApcMin ) using Sleeping Beauty (SB ) transposon-mediated mutagenesis.ApcMin SB-mutagenized mice developed three times as many polyps as mice with theApcMin allele alone. Analysis of transposon common insertion sites (CIS) identified theApc locus as a major target ofSB -induced mutagenesis, suggesting thatSB insertions provide an efficient route to biallelicApc inactivation. We also identified an additional 32 CIS genes/loci that may represent modifiers of theApcMin 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.