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Application of reagent for inhibiting expression of SMC1B gene

A technology of gene expression and reagents, applied in the field of medical engineering, can solve the problems of reports of no function of SMC1B gene, no function of SMC1B gene, etc.

Active Publication Date: 2018-04-24
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, there is no report that the SMC1B gene plays a role in HBsAg, let alone a report that the SMC1B gene plays a role in truncated HBsAg

Method used

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  • Application of reagent for inhibiting expression of SMC1B gene
  • Application of reagent for inhibiting expression of SMC1B gene
  • Application of reagent for inhibiting expression of SMC1B gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 The method of the present invention knocks out the SMC1B gene and verification

[0041] 1. Experimental materials

[0042] (1) Plasmids, cell lines and experimental animals

[0043] 1. Plasmids and cell lines

[0044] (1) Perform related experiments using the L02 cell line stably transfected with replicating HBV recombinant plasmids. In vitro transfection of pHBV4.1 plasmid into cells or mice can achieve the complete transcription, replication and expression of HBV genes:

[0045] pHBV4.1 (express wild-type HBsAg)

[0046] pHBV4.1-sW172* (Expressing truncated HBsAg)

[0047] pHBV4.1-sW172L (Expression replacement HBsAg)

[0048] (2) Use CRISPR / Cas9 gene knockout system to complete SMC1 gene knockout (Santa Cruz Biotech, USA), see the structure of CRISPR / Cas9 and HDR plasmid figure 1 .

[0049] A: SMC1CRISPR / Cas9KO Plasmid(h) Plasmid: SMC1 knockout plasmid, using three different sgRNA sequences as guide RNA to knock out the target gene.

[0050] B: SMC1HDR co-transformation p...

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Abstract

The invention provides an application of a reagent for inhibiting expression of an SMC1B gene in preparing medicines for preventing and / or treating canceration of hepatitis B patients. The invention also provides a kit for knocking out the SMC1B gene and a method for preparing an SMC1B gene knockout cell model. The inventor, after knocking out the SMC1B gene by virtue of a specific sgRNA sequence,can reduce the canceration of HBV, and especially can reduce the canceration of a BV rtA181T / sW172* mutant strain quite obviously; therefore, important theoretical and practical significance can be achieved on clinical prevention and treatment of HBV related diseases.

Description

Technical field [0001] The invention belongs to the technical field of medical engineering, and specifically relates to the use of a reagent for inhibiting the expression of SMC1B gene. Background technique [0002] At present, at least 250 million people in the world are infected with hepatitis B virus (HBV). Persistent HBV infection can lead to the occurrence of chronic hepatitis, cirrhosis and primary liver cancer, resulting in approximately 650,000 deaths each year. There are two main categories of drugs approved by the State Food and Drug Administration for anti-HBV treatment: IFN and nucleoside (acid) analogs [nucleos(t)ideanalogs, NAs]. Among them, the adverse reactions during IFN treatment are serious, and only a small number of people can tolerate IFN treatment; while NAs has fewer adverse reactions, convenient administration, and strong effects, so it has good clinical application value. [0003] However, with the prolongation of NAs antiviral drugs, the problem of resis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K48/00A61P35/00C12N15/90C12N9/22C12N5/10
CPCA61K45/00A61K48/005C12N5/067C12N9/22C12N15/907C12N2510/00
Inventor 唐红周凌云陈恩强杜凌遥白浪汪梦兰
Owner WEST CHINA HOSPITAL SICHUAN UNIV