Assessment of Fecundity and Germ Line Transmission in Two Transgenic Pig Lines Produced by Sleeping Beauty Transposition

Genes

Wiebke Garrels²  Stephanie Holler²  Nicole Cleve²  Heiner Niemann²  Zoltan Ivics¹ 

[1] Paul-Ehrlich-Institute, Paul-Ehrlich-Straße 51-59, 63225 Langen, Germany; E-Mail:;Friedrich-Loeffler-Institut, Institute of Farm Animal Genetics, Höltystraße 10, 31535 Neustadt, Germany; E-Mails:

关键词: transgenic animal;    germ line transmission;    active transgenesis;    hyperactive transposase;    humanized pig model;    livestock;    cytoplasmic plasmid injection;    gene silencing;    permissive locus;   

DOI  :  10.3390/genes3040615

来源: mdpi

【 摘 要 】

Recently, we described a simplified injection method for producing transgenic pigs using a non-autonomous Sleeping Beauty transposon system. The founder animals showed ubiquitous expression of the Venus fluorophore in almost all cell types. To assess, whether expression of the reporter fluorophore affects animal welfare or fecundity, we analyzed reproductive parameters of two founder boars, germ line transmission, and organ and cell specific transgene expression in animals of the F1 and F2 generation. Molecular analysis of ejaculated sperm cells suggested three monomeric integrations of the Venus transposon in both founders. To test germ line transmission of the three monomeric transposon integrations, wild-type sows were artificially inseminated. The offspring were nursed to sexual maturity and hemizygous lines were established. A clear segregation of the monomeric transposons following the Mendelian rules was observed in the F1 and F2 offspring. Apparently, almost all somatic cells, as well as oocytes and spermatozoa, expressed the Venus fluorophore at cell-type specific levels. No detrimental effects of Venus expression on animal health or fecundity were found. Importantly, all hemizygous lines expressed the fluorophore in comparable levels, and no case of transgene silencing or variegated expression was found after germ line transmission, suggesting that the insertions occurred at transcriptionally permissive loci. The results show that Sleeping Beauty transposase-catalyzed transposition is a promising approach for stable genetic modification of the pig genome.

【 授权许可】

CC BY   
© 2012 by the authors; licensee MDPI, Basel, Switzerland.