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RNA interference sequence aiming at hepatocyte nuclear factor 4a and applications thereof

A technology of hepatocyte nuclear factor and RNA interference, which is applied in the field of RNA interference sequence specifically targeting hepatocyte nuclear factor 4a, achieves good application prospects and inhibits HBV transcription and replication

Active Publication Date: 2012-01-04
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on RNAi targeting HNF4α at present, and there is a need to develop new effective drugs for HBV treatment in this field

Method used

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  • RNA interference sequence aiming at hepatocyte nuclear factor 4a and applications thereof
  • RNA interference sequence aiming at hepatocyte nuclear factor 4a and applications thereof
  • RNA interference sequence aiming at hepatocyte nuclear factor 4a and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 Obtaining of interfering RNA sequence of the present invention and its anti-HBV efficacy

[0028] Materials and methods

[0029] 1. Plasmid construction

[0030] The present invention uses human HNF4α mRNA (GenBank accession number: X87870) as a template to design shRNA target sequences. Four pairs of interference sequences targeting HNF4α (shRNA1, shRNA2, shRNA3, shRNA4, see Table 1) were designed, and fragments of the conserved sequence of the open reading frame were designed (each 55bp, synthesized by Invitrogen, and the designs are shown in SEQ ID No.4, SEQ ID No.5) and cloned into pSilencer 4.1-CMV puro vector between the BamHI and HindIII sites respectively.

[0031] The hepatocyte nuclear factor 4a RNA interference sequence designed in Table 1

[0032]

[0033] The sense strand (Top Strand Oligonucleotide Template) of the expression framework is shown in SEQ ID No.4:

[0034] 5′-GATCC CCACArGTACTCCTGCAGA TTCAAGAGA TCTGCAGGAGTACATGTGG TT A...

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Abstract

The invention belongs to the field of hepatitis B prevention and treatment, and in particular relates to a hepatocyte nuclear factor 4a specificity short hairpin RNA with an inhibition effect for hepatitis B virus replication and applications thereof. The technical problem to be solved by the invention is to provide a novel effective choice for HBV resistant treatment. The technical scheme of the technical problem solved by the invention is to provide a specificity short hairpin RNA aiming at the hepatocyte nuclear factor 4a and a carrier capable of expressing the sequence. The shRNA sequence and the expression carrier thereof provided by the invention are testified to be able to inhibit internal HBV transcription and copy effectively and have no obvious side effects, and provide a novel effective choice for HBV resistance, and have favorable application prospects in preparing HBV-resistant medicaments.

Description

technical field [0001] The invention belongs to the field of prevention and treatment of hepatitis B, and in particular relates to an RNA interference sequence specifically targeting hepatocyte nuclear factor 4a and its application. Background technique [0002] Hepatitis B virus (HBV) infection is a serious global health problem. It is estimated that approximately 350 million people are chronic HBV carriers who are prone to develop liver failure, cirrhosis, and / or hepatocellular carcinoma (HCC) [1-3]. Recent studies have shown that active HBV replication is closely related to disease progression, especially sustained high levels of serum HBV DNA increase the risk of liver cirrhosis and HCC[4-7]. Currently, there are two classes of approved anti-HBV drugs, including interferon and nucleoside / nucleotide analogues, and it has been observed that controlling HBV replication with antiviral drugs reduces the risk of developing cirrhosis and liver cancer. However, long-term treat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63A61K48/00A61P1/16A61P31/20
Inventor 唐红何芳刘丽周陶友白浪陈恩强刘聪程星
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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