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Regulation of porcine endogenous retrovirus by dual LTR1+2 (Long Terminal Region) miRNA in primary porcine kidney cells

Journal of Veterinary Science 2019년 20권 5호 p.50 ~ 50
 ( Chung Hee-Chun ) - Seoul National University College of Veterinary Medicine

 ( Nguyen Van-Giap ) - Vietnam National University of Agriculture Faculty of Veterinary Medicine Department of Veterinary Microbiology-Infectious Diseases
 ( Moon Hyung-Joon ) - Green Cross Veterinary Products Research Unit
 ( Park Yong-Ho ) - Seoul National University College of Veterinary Medicine Department of Veterinary Microbiology
 ( Park Bong-Kyun ) - Seoul National University College of Veterinary Medicine

Abstract


Porcine endogenous retroviruses (PERVs) integrate into germline DNA as proviral genome that enables vertical transmission from parents to their offspring. The provirus usually survives as part of the host genome rather than as an infectious agent, but may become pathogenic if it crosses species barriers. Therefore, replication-competent PERV should be controlled through selective breeding or knockout technologies. Two microRNAs (miRNAs), dual LTR1 and LTR2, were selected to inhibit the expression of PERV in primary porcine kidney cells. The inhibition efficiency of the miRNAs was compared based on their inhibition of different PERV regions, specifically long terminal repeats (LTRs), gag, pol, and env. Gene expression was quantified using real-time polymerase chain reaction and the C-type reverse transcriptase (RT) activity was determined. The messenger RNA (mRNA) expression of the PERV LTR and env regions was determined in HeLa cells co-cultured with primary porcine kidney cells. The mRNA expression of the LTR, gag, pol, and env regions of PERV was dramatically inhibited by dual miRNA from 24 to 144 h after transfection, with the highest inhibition observed for the LTR and pol regions at 120 h. Additionally, the RT activity of PERV in the co-culture experiment of porcine and human cells was reduced by 84.4% at the sixth passage. The dual LTR 1+2 miRNA efficiently silences PERV in primary porcine kidney cells.

키워드

Porcine endogenous retrovirus; inhibition; miRNA; long terminal region; primary porcine kidney cell
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