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In vivo site-specific integration of transgene in silkworm via PhiC31 integrase-mediated cassette exchange.

Abstract：

:Current techniques for genetic engineering of the silkworm Bombyx mori genome utilize transposable elements, which result in positional effects and insertional mutagenesis through random insertion of exogenous DNA. New methods for introducing transgenes at specific positions are therefore needed to overcome the limitations of transposon-based strategies. Although site-specific recombination systems have proven powerful tools for genome manipulation in many organisms, their use has not yet been well established for the integration of transgenes in the silkworm. We describe a method for integrating target genes at pre-defined chromosomal sites in the silkworm via phiC31/att site-specific recombination system-mediated cassette exchange. Successful recombinase-mediated cassette exchange (RMCE) was observed in the two transgenic target strains with an estimated transformation efficiency of 3.84-7.01%. Our results suggest that RMCE events between chromosomal attP/attP target sites and incoming attB/attB sites were more frequent than those in the reciprocal direction. This is the first report of in vivo RMCE via phiC31 integrase in the silkworm, and thus represents a key step toward establishing genome manipulation technologies in silkworms and other lepidopteran species.

journal_name

Insect Biochem Mol Biol

journal_title

Insect biochemistry and molecular biology

authors

Long D,Zhao A,Xu L,Lu W,Guo Q,Zhang Y,Xiang Z

doi

10.1016/j.ibmb.2013.08.001

subject

Has Abstract

pub_date

2013-11-01 00:00:00

pages

997-1008

issue

11

eissn

0965-1748

issn

1879-0240

pii

S0965-1748(13)00148-3

journal_volume

43

pub_type

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