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- Title
Targeted rearrangement of a chromosomal repeat sequence by transfection of a homologous DNA sequence using purified integrase.
- Authors
Tanaka, A. S.; Komuro, K.
- Abstract
Using a liposomal transfection with purified bovine leukemia virus (BLV) integrase, we observed an efficient DNA rearrangement of a chromosomal repeat sequence and targeted integration of a part of the transfected plasmid. The BLV integrase recognition sequence (IRS) including the 3'end of the BLV LTR U5, one of the sites cleaved by the integrase, was essential for the DNA rearrangement, and a sequence homologous to the chromosomal DNA neighboring the repeat target site had to be placed downstream of the IRS on the transfected plasmid. The pSV2neo DNA, including the pBR322 sequence preintegrated into L929 cells (primary transfectants), was rearranged by a secondary transfection of a pBR322-based hygromycin-resistance plasmid carrying the IRS. We present a model to explain the chromosomal DNA rearrangement of the primary clones through a homologous recombination-like reaction and amplification of the neighboring sequences.Gene Therapy (2005) 12, 783-794. doi:10.1038/sj.gt.3302458 Published online 17 February 2005
- Subjects
VIRUS-induced enzymes; BOVINE leukemia virus; VIRAL genetics; GENE transfection; NUCLEOTIDE sequence; GENE amplification
- Publication
Gene Therapy, 2005, Vol 12, Issue 9, p783
- ISSN
0969-7128
- Publication type
Article
- DOI
10.1038/sj.gt.3302458