TAL effectors mediate high-efficiency transposition of the piggyBac transposon in silkworm Bombyx mori L

The piggyBac ( PB ) transposon is one of the most useful transposable elements and has been successfully used for genetic manipulation in more than a dozen species. However, the efficiency of PB -mediated transposition is still insufficient for many purposes. Here, we present a strategy to enhance t...

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Bibliographic Details
Published in:Scientific reports 2015-11, Vol.5 (1), p.17172-17172, Article 17172
Main Authors: Ye, Lupeng, You, Zhengying, Qian, Qiujie, Zhang, Yuyu, Che, Jiaqian, Song, Jia, Zhong, Boxiong
Format: Article
Language:English
Subjects:
42
42/44
42/70
631/1647/1513
631/337/2569
631/61/2300/1514
Animals
Animals, Genetically Modified
Bombyx - genetics
DNA Transposable Elements - genetics
Embryo, Nonmammalian - metabolism
Gene Transfer Techniques
Green Fluorescent Proteins - metabolism
Humanities and Social Sciences
multidisciplinary
Mutagenesis, Insertional
Plasmids - metabolism
Science
Trans-Activators - metabolism
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Summary:The piggyBac ( PB ) transposon is one of the most useful transposable elements and has been successfully used for genetic manipulation in more than a dozen species. However, the efficiency of PB -mediated transposition is still insufficient for many purposes. Here, we present a strategy to enhance transposition efficiency using a fusion of transcription activator-like effector (TALE) and the PB transposase ( PBase ). The results demonstrate that the TALE- PBase fusion protein which is engineered in this study can produce a significantly improved stable transposition efficiency of up to 63.9%, which is at least 7 times higher than the current transposition efficiency in silkworm. Moreover, the average number of transgene-positive individuals increased up to 5.7-fold, with each positive brood containing an average of 18.1 transgenic silkworms. Finally, we demonstrate that TALE- PBase fusion-mediated PB transposition presents a new insertional preference compared with original insertional preference. This method shows a great potential and value for insertional therapy of many genetic diseases. In conclusion, this new and powerful transposition technology will efficiently promote genetic manipulation studies in both invertebrates and vertebrates.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep17172