title: TAL effectors mediate high-efficiency transposition of the piggyBac transposon in silkworm Bombyx mori L.
abstract: 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.
authors.affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
authors.forename: Lupeng
authors.initials: L
authors.lastname: Ye
authors.affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
authors.forename: Zhengying
authors.initials: Z
authors.lastname: You
authors.affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
authors.forename: Qiujie
authors.initials: Q
authors.lastname: Qian
authors.affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
authors.forename: Yuyu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
authors.forename: Jiaqian
authors.initials: J
authors.lastname: Che
authors.affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
authors.forename: Jia
authors.initials: J
authors.lastname: Song
authors.affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
authors.forename: Boxiong
authors.initials: B
authors.lastname: Zhong
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D015534:Trans-Activators; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins