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- Title
Inhibition of porcine endogenous retrovirus in PK15 cell line by efficient multitargeting RNA interference.
- Authors
Chung, Hee-Chun; Nguyen, Van-Giap; Moon, Hyoung-Joon; Kim, Hye-Kwon; Park, Seong-Jun; Lee, Jee-Hoon; Choi, Min-Gyung; Kim, A-Reum; Park, Bong-Kyun
- Abstract
To effectively suppress porcine endogenous retroviruses ( PERV)s, RNAi technique was utilized. RNAi is the up-to-date skill for gene knockdown which simultaneously multitargets both gag and pol genes critical for replication of PERVs. Previously, two of the most effective si RNAs (gag2, pol2) were found to reduce the expression of PERVs. Concurrent treatment of these two si RNAs (gag2+pol2) showed knockdown efficiency of up to 88% compared to negative control. However, despite the high initial knockdown efficiency 48 h after transfection caused by si RNA, it may only be a transient effect of suppressing PERVs. The multitargeting vector was designed, containing both gag and pol genes and making use of POL II mi R Expression Vector, which allowed for persistent and multiple targeting. This is the latest sh RNA system technique expressing and targeting like mi RNA. Through antibiotics resistance characteristics utilizing this vector, mi RNA-transfected PK15 cells (gag2-pol2) were selected during 10 days. An 88.1% reduction in the level of mRNA expression was found. In addition, we performed RT-activity analysis and fluorescence in situ hybridization assay, and it demonstrated the highest knockdown efficiency in multitargeting (gag2+pol2) mi RNA group. Therefore, according to the results above, gene knockdown system (si RNA and sh RNA) through multitargeting strategy could effectively inhibit PERVs.
- Subjects
PORCINE somatotropin; RETROVIRUSES; RNA interference; DNA replication; SMALL interfering RNA; GENE expression; DRUG resistance
- Publication
Transplant International, 2014, Vol 27, Issue 1, p96
- ISSN
0934-0874
- Publication type
Article
- DOI
10.1111/tri.12219