Identification and targeting of the pig Rosa26 locus

Author: Huiming Ju 1, 2,¶, Yuefei Yang , Lijing Bai 2, Lei Huang 2, Xin Jiang 1, Yulian Mu 2* and Kui Li 2*
Received 2 January 2015, accepted 30 March 2015.
Abstract

The Rosa26 locus in mice is an ubiquitously expressed gene that has a high homologous recombination efficiency. We have investigated the orthologous gene in pig, Sus scrofa. The mouse-transcribed Rosa26 (mRosa26) sequence was used to screen the pig genome database to identify homologous sequences. Four different pig Rosa26 (pRosa26) promoter sequences were cloned into a luciferase expression reporter vector and a 409-bp sequence was shown to be a core promoter. Zinc finger nucleases were designed based on the pRosa26 exon 1 sequence and used to mediate gene targeting in the pig PK15 cell line. Targeted cell lines were identified and pRosa26 gene targeting verified. β-Galactosidase staining of targeted clones indicated that the Rosa26 promoter was functional. Cell cycle and apoptosis indices demonstrated no significant differences between targeted and control cells. This study confirmed the existence of pig Rosa26 and identified it as a suitable locus for transgenic manipulation.

Journal: Food, Agriculture and Environment (JFAE)
Online ISSN: 1459-0263Year: 2015, Vol. 13, Issue 2, pages 149-155.Publisher: WFL.


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