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Genes 2018, 9(12), 630; https://doi.org/10.3390/genes9120630

Enhancer Trapping and Annotation in Zebrafish Mediated with Sleeping Beauty, piggyBac and Tol2 Transposons

1
Institute of Mobilome and Genome, College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China
2
Institute of Cancer and Genomic Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Received: 5 September 2018 / Revised: 7 December 2018 / Accepted: 10 December 2018 / Published: 13 December 2018
(This article belongs to the Section Molecular Genetics and Genomics)
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Abstract

Although transposon-mediated enhancer trapping (ET) is successfully applied in diverse models, the efficiency of various transposon systems varies significantly, and little information is available regarding efficiency of enhancer trapping by various transposons in zebrafish. Most potential enhancers (Ens) still lack evidence of actual En activity. Here, we compared the differences in ET efficiency between sleeping beauty (SB), piggyBac (PB) and Tol2 transposons. Tol2 represented the highest germline transfer efficiencies at 55.56% (NF0 = 165), followed by SB (38.36%, NF0 = 151) and PB (32.65%, NF0 = 149). ET lines generated by the Tol2 transposon tended to produce offspring with a single expression pattern per line, while PB and SB tended to generate embryos with multiple expression patterns. In our tests, 10 putative Ens (En1–10) were identified by splinkerette PCR and comparative genomic analysis. Combining the GFP expression profiles and mRNA expression patterns revealed that En1 and En2 may be involved in regulation of the expression of dlx1a and dlx2a, while En6 may be involved in regulation of the expression of line TK4 transgene and rps26, and En7 may be involved in the regulation of the expression of wnt1 and wnt10b. Most identified Ens were found to be transcribed in zebrafish embryos, and their regulatory function may involve eRNAs. View Full-Text
Keywords: enhancer trapping; enhancer; Sleeping Beauty; piggyBac; Tol2; zebrafish enhancer trapping; enhancer; Sleeping Beauty; piggyBac; Tol2; zebrafish
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Shen, D.; Xue, S.; Chan, S.; Sang, Y.; Wang, S.; Wang, Y.; Chen, C.; Gao, B.; Mueller, F.; Song, C. Enhancer Trapping and Annotation in Zebrafish Mediated with Sleeping Beauty, piggyBac and Tol2 Transposons. Genes 2018, 9, 630.

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