title: Short hairpin RNAs with a 2- or 3-base mismatch inhibit HBV expression and replication in HepG2 cells.
abstract: OBJECTIVE
  To assess the functions of mismatched short hairpin RNAs (shRNAs) that inhibit replication and the expression of hepatitis B virus (HBV), two shRNAs possessing a 2- or 3-base mismatch that targeted HBV were studied.
  
  
  METHODS
  shRNAs and pHY106-HBV were cotransfected into HepG2 cells. The culture supernatants were collected and used in hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) assays. The levels of HBsAg and HBcAg mRNA were detected by reverse-transcriptase PCR (RT-PCR). HBV DNA replication intermediates were extracted for Southern blot hybridization.
  
  
  RESULTS
  The results demonstrate that mismatched shRNA-458 and shRNA-635 can significantly inhibit HBsAg and HBeAg protein expression, and the maximal inhibition ratio for both proteins was found at 72 h after cotransfection: 80 and 50 %, respectively. Similar inhibitory effects were found on HBsAg and HBcAg mRNA levels and HBV DNA replication intermediates at 72 h after cotransfection, and the inhibition ratio was found to be approximately 70 and 90 %, respectively.
  
  
  CONCLUSIONS
  Despite the 2- or 3-base mismatch between the shRNAs and the HBV target sequences, shRNA-458 and shRNA-635 exerted a significant inhibitory effect on HBsAg and HBeAg expression and HBV replication. This indicates that mismatched shRNAs could be a promising therapy for HBV.
authors.affiliation: Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, No. 8 Jingshun East Street, Chaoyang District, Beijing, 100015, China.|Department of Infectious Diseases, The Second Affiliated Hospital of Harbin Medical University, No. 246 Xuefu Road, Harbin, 150086, Heilongjiang, China.
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Li
authors.affiliation: Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, No. 8 Jingshun East Street, Chaoyang District, Beijing, 100015, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Hong
authors.affiliation: Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, No. 8 Jingshun East Street, Chaoyang District, Beijing, 100015, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, No. 8 Jingshun East Street, Chaoyang District, Beijing, 100015, China.
authors.forename: Shunai
authors.initials: S
authors.lastname: Liu
authors.affiliation: Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, No. 8 Jingshun East Street, Chaoyang District, Beijing, 100015, China.
authors.forename: Hongshan
authors.initials: H
authors.lastname: Wei
authors.affiliation: Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, No. 8 Jingshun East Street, Chaoyang District, Beijing, 100015, China. jun.cheng.ditanhospital@gmail.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
keywords: HBV; Mismatch; RNA; Short hairpin