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KMID : 1023620060300030181
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Reproductive and Developmental Biology
2006 Volume.30 No. 3 p.181 ~ p.187
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Inhibition of Porcine Endogenous Retrovirus Expression by RNA Interference
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Lee Hyun-Na
Koo Bon-Chul
Kwon Mo-Seon
Kim Teo-An
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Abstract
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In recent years the number of patients waiting for organ transplantation has greatly outpaced the supply of human organs available, which leads to a renewed interest in pig-to-human xenotransplantation as an alternative. However, one of the biggest barriers in the xenotransplantation is presence of porcine endogenous retroviruses(PERV) that can infect human cells. In this study, to present a possible solution for this problem we tried to inhibit expression of PERVs using shRNAs(short hairpin RNA) at the level of RNA synthesis and virus release. The shRNA targeting the sequence of PERV A, B type was cloned into pSIREN-RetroQ vector under the control of polymerase-III U6-RNA gene promoter. Quantitative real-time PCR was performed to detect my alterations in mRNA production of PERV A, B targeted by the shRNA in each done. Depending on the target sequence of the shRNA, the transcription of PERV was decreased to as much as 4% and the number of progeny viruses was reduced to less than 1/200,000. Transgenic pigs producing such shRNAs may result in a highly reduced PERV expression in cells and organs, which is a prerequisite for safe xenotransplantations.
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KEYWORD
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Porcine endogenous retrovirus, RNA interference, Short hairpin RNA, Xenotransplantation
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