Novel Chimeric Vectors Harboring Hepatitis B Viral Promoter and Reporter Gene Demonstrated Liver-Specific Significance

Abstract

Aim: Expression of EGFP was investigated to ascertain the strength and specificity of CMV, U6 and hepatitis B virus (HBV) core promoters in hepatic and non-hepatic cells. Materials & methods: pSilencer-2.1 plasmid vector is known for siRNA-based inhibition. To achieve target-specific correction of disease-causing genes, pSilencer–GFP was constructed. For liver specific expression of therapeutic genes, endogenous U6 promoter of pSilencer-2.1 was replaced with HBV core promoter and ubiquitously active CMV promoter. Results: Transfection results showed that GFP expression under the control of HBV core promoter was higher in hepatic Hep3B than non-hepatic HEK293T cells. Conclusion: HBV core promoter could lead to specific expression in hepatocytes, which might be used in gene therapy of liver diseases as well as for siRNA-based therapeutic strategies.

Keywords:

• CMV promoter
• gene therapy
• hepatitis B viral promoter
• pSilencer-2.1 vector
• U6 promoter

Author contributions

A Anwer and SN Kazim conceived the idea and planned the study. A Anwer conducted the major experiments, analyzed the data and prepared a major part of the manuscript. A Anwer, S Khan, F Amir, M Afroz and SA Azam made contributions to the experiments as well as in critically reviewing the manuscript. SA Bhat, SK Hasan and R Waseem made their contributions with both the experiments and data analysis. S Parveen, A Islam and SN Kazim gave finishing touches to the manuscript.

Financial & competing interests disclosure

Financial support from DST WOS-A grant no. SR/WOS-A/LS-255/2010(G) was granted to M Afroz and DBT project grant no. BT/PR10740/MED/29/79/2008 was granted to SN Kasim. The first author A Anwer acknowledges receipt of the Senior Research Fellowship (SRF), Indian Council of Medical Research (ICMR), Government of India, for the fellowship support under grant no. VIR/Fellowship/34/2019-ECD-I. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.