Use of mariner transposases for one-step delivery and integration of DNA in prokaryotes and eukaryotes by transfection.

ABSTRACT: Delivery of DNA to cells and its subsequent integration into the host genome is a fundamental task in molecular biology, biotechnology and gene therapy. Here we describe an IP-free one-step method that enables stable genome integration into either prokaryotic or eukaryotic cells. A synthetic mariner transposon is generated by flanking a DNA sequence with short inverted repeats. When purified recombinant Mos1 or Mboumar-9 transposase is co-transfected with transposon-containing plasmid DNA, it penetrates prokaryotic or eukaryotic cells and integrates the target DNA into the genome. In vivo integrations by purified transposase can be achieved by electroporation, chemical transfection or Lipofection of the transposase:DNA mixture, in contrast to other published transposon-based protocols whic

SUBMITTER: Trubitsyna M 

PROVIDER: S-EPMC5449632 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature