Somatic integration of an oncogene-harboring Sleeping Beauty transposon models liver tumor development in the mouse.
The Sleeping Beauty (SB) transposon system can integrate foreign sequences of DNA in the genome of mouse somatic cells eliciting long-term expression in vivo. This technology holds great promise for human gene therapy as a nonviral technology to deliver therapeutic genes. SB also provides a means to study the effects of defined genetic elements, such as oncogenes, on somatic cells in mice. Here, we test the ability of the SB transposon system to facilitate somatic integration of a transposon containing an activated NRAS oncogene in mouse hepatocytes to elicit tumor formation. NRAS oncogene-driven tumors developed when such vectors were delivered to the livers of p19Arf-null or heterozygous mice. Delivery of the NRAS transposon cooperates with Arf loss to cause carcinomas of hepatocellular or biliary origin. These tumors allowed characterization of transposon integration and expression at the single-cell level, revealing robust NRAS expression and both transposase-mediated and random insertion of delivered vectors. Random integration and expression of the SB transposase plasmid was also observed in one instance. In addition, studies using effector loop mutants of activated NRAS provide evidence that mitogen-activated protein kinase activation alone cannot efficiently induce liver carcinomas. This system can be used to rapidly model tumors caused by defined genetic changes.
Carlson
Corey M
CM
The Arnold and Mabel Beckman Center for Transposon Research, Institute of Human Genetics, Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN 55455, USA.
Frandsen
Joel L
JL
Kirchhof
Nicole
N
McIvor
R Scott
RS
Largaespada
David A
DA
D000818:Animals; D002277:Carcinoma / Q000209:etiology / Q000235:genetics / Q000473:pathology; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004251:DNA Transposable Elements; D004195:Disease Models, Animal; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008113:Liver Neoplasms / Q000209:etiology* / Q000235:genetics / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D013154:Spleen / Q000473:pathology; D019895:Transposases / Q000235:genetics* / Q000378:metabolism; D025803:Tumor Suppressor Protein p14ARF / Q000235:genetics
C417743:Cdkn2a protein, mouse; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004251:DNA Transposable Elements; D025803:Tumor Suppressor Protein p14ARF; D019895:Transposases; C494471:sleeping beauty transposase, human