Molecular Reconstruction of Sleeping Beauty, a Tc1-like Transposon From Fish, and Its Transposition in Human Cells

Cell. 1997 Nov 14;91(4):501-10. doi: 10.1016/s0092-8674(00)80436-5.

Abstract

Members of the Tc1/mariner superfamily of transposons isolated from fish appear to be transpositionally inactive due to the accumulation of mutations. Molecular phylogenetic data were used to construct a synthetic transposon, Sleeping Beauty, which could be identical or equivalent to an ancient element that dispersed in fish genomes in part by horizontal transmission between species. A consensus sequence of a transposase gene of the salmonid subfamily of elements was engineered by eliminating the inactivating mutations. Sleeping Beauty transposase binds to the inverted repeats of salmonid transposons in a substrate-specific manner, and it mediates precise cut-and-paste transposition in fish as well as in mouse and human cells. Sleeping Beauty is an active DNA-transposon system from vertebrates for genetic transformation and insertional mutagenesis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Carps
  • Cell Line
  • DNA / metabolism
  • DNA Transposable Elements / genetics*
  • DNA, Recombinant / genetics
  • Enzyme Activation
  • Humans
  • Mice
  • Molecular Sequence Data
  • Recombinant Fusion Proteins
  • Salmonidae / genetics*
  • Transfection
  • Transposases / chemistry
  • Transposases / genetics*
  • Transposases / metabolism*

Substances

  • DNA Transposable Elements
  • DNA, Recombinant
  • Recombinant Fusion Proteins
  • DNA
  • Transposases