Transposon-mediated mutagenesis in somatic cells: identification of transposon-genomic DNA junctions

Methods Mol Biol. 2008;435:95-108. doi: 10.1007/978-1-59745-232-8_7.

Abstract

Understanding the genetic basis for tumor formation is crucial for treating cancer. Forward genetic screens using insertional mutagenesis technologies have identified many important tumor suppressor genes and oncogenes in mouse models of human cancer. Traditionally, retroviruses have been used for this purpose, allowing the identification of genes that can cause various forms of leukemia or lymphoma with murine leukemia viruses or mammary cancer with mouse mammary tumor viruses. Recently, the Sleeping Beauty transposon system has emerged as a tool for cancer gene discovery in mouse models of human cancer. Transposons mobilized in the mouse soma can insertionally mutate cancer genes, and the transposon itself serves as a molecular "tag," which facilitates candidate cancer gene identification. We provide an overview of some general issues related to use of Sleeping Beauty for cancer genetic studies and present here the polymerase chain reaction-based method for cloning transposon-tagged sequences from tumors.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers / genetics
  • DNA Transposable Elements / genetics*
  • Genes, Tumor Suppressor
  • Humans
  • Mice
  • Mutagenesis, Insertional / methods*
  • Neoplasms, Experimental / etiology
  • Neoplasms, Experimental / genetics
  • Oncogenes
  • Polymerase Chain Reaction
  • Transposases / genetics

Substances

  • DNA Primers
  • DNA Transposable Elements
  • Transposases
  • sleeping beauty transposase, human