NDLI: Highly efficient and inducible DNA excision in transgenic silkworms using the FLP/FRT site-specific recombination system

Content Provider	Springer Nature Link
Author	Long, Dingpei Lu, Weijian Hao, Zhanzhang Xiang, Zhonghuai Zhao, Aichun
Copyright Year	2016
Abstract	Efficient and inducible recombinase-mediated DNA excision is an optimal technology for automatically deleting unwanted DNA sequences, including selection marker genes. However, this methodology has yet to be established in transgenic silkworms. To achieve efficient and inducible FLP recombinase-mediated DNA excision in transgenic silkworms, one transgenic target strain (TTS) containing an FRT-flanked silkworm cytoplasmic actin 3 gene promoter (A3)-enhanced green fluorescent protein (EGFP) expression cassette, as well as two different types of FLP recombinase expression helper strains were generated. Then, the FLP recombinase was introduced into the TTS silkworms by pre-blastoderm microinjection and sexual hybridization. Successful recombinase-mediated deletion of the A3-EGFP expression cassette was observed in the offspring of the TTS, and the excision efficiencies of the FLP expression vector and FLP mRNA pre-blastoderm microinjection were 2.38 and 13.3 %, respectively. The excision efficiencies resulting from hybridization between the TTS and the helper strain that contained a heat shock protein 70 (Hsp70)-FLP expression cassette ranged from 32.14 to 36.67 % after heat shock treatment, while the excision efficiencies resulting from hybridization between the TTS and the helper strain containing the A3-FLP expression cassette ranged from 97.01 to 100 %. These results demonstrate that the FLP/FRT system can be used to achieve highly efficient and inducible post-integration excision of unwanted DNA sequences in transgenic silkworms in vivo. Our present study will facilitate the development and application of the FLP/FRT system for the functional analysis of unknown genes, and establish the safety of transgenic technologies in the silkworm and other lepidopteran species.
Starting Page	795
Ending Page	811
Page Count	17
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	25
Issue Number	6
e-ISSN	15739368
Language	English
Publisher	Springer International Publishing
Publisher Date	2016-06-22
Publisher Place	Cham
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Bombyx mori FLP/FRT system Safety of transgenic technologies Site-specific excision Transgenic silkworms Animal Genetics and Genomics Plant Genetics & Genomics Transgenics Biomedical Engineering/Biotechnology Genetic Engineering Molecular Medicine
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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