Open AccessFLP Recombinase-Mediated Site-Specific Recombination in Silkworm, Bombyx mori
Author(s) -
Dan Long,
Aichun Zhao,
Xue-Jiao Chen,
Yang Zhang,
Weijian Lu,
Qing Guo,
Alfred M. Handler,
Xiang Zhang
Publication year - 2012
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0040150
Subject(s) - site specific recombination , biology , genome , flp frt recombination , genetics , recombinase , transgene , bombyx , gene , bombyx mori , recombineering , genome engineering , gene targeting , computational biology , genome editing , recombination , genetic recombination
A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/ FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/ FRT system. We conclude that the FLP/ FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/ FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species.