Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference.

Abstract:

:Bovine viral diarrhea virus (BVDV) should be a ubiquitous viral pathogen to the cattle and sheep industry. This pathogen is responsible for severe economic losses. We previously showed that plasmid-mediated dual short hairpin RNA (shRNA) efficiently inhibit BVDV replication in bovine kidney epithelial (MDBK) cells. In this study, we delivered the dual shRNA system to sheep fibroblasts and generated transgenic cell colonies. These transgenic fibroblasts were further used for somatic cell nuclear transfer (SCNT). Three lambs were born at full term, but perished soon after birth. Integration of shRNA into the genome of cloned sheep was confirmed by PCR and expression of shRNA in transgenic sheep was confirmed by real-time PCR. Kidney epithelial cells were isolated from transgenic sheep and challenged with multiple BVDV subgenotypes (BVDV-1a, BVDV-1b and BVDV-1c). The dual shRNA expressed in transgenic kidney epithelial cells significantly inhibited BVDV replication in a cross-resistance manner. Our results showed that transgenic RNAi might be a useful tool for preparation of transgenic animals with increased resistance to BVDV.

journal_name

J Virol Methods

authors

Ni W,Qiao J,Ma Q,Wang J,Wang D,Zhao X,Cao Y,Li Q,Hu S,Chen C

doi

10.1016/j.jviromet.2015.03.014

subject

Has Abstract

pub_date

2015-06-15 00:00:00

pages

66-70

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(15)00104-4

journal_volume

218

pub_type

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