System for inducing formation of HBV cccDNA and construction method thereof

A construction method and fragment technology, which are applied in the field of inducing the formation of HBV cccDNA, can solve the problems of harsh infection conditions, low sensitivity, high cost, etc., and achieve the effect of simple biological background.
CN110669735APending Publication Date: 2020-01-10GUANGZHOU EIGHTH PEOPLES HOSPITAL
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Patent Information

Authority / Receiving Office
CN ยท China
Current Assignee / Owner
GUANGZHOU EIGHTH PEOPLES HOSPITAL
Publication Date
2020-01-10

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Abstract

The invention discloses a system for inducing formation of HBV cccDNA and a construction method thereof. According to the invention, a stable cell line is constructed by integrating a Phic31 recombinase gene; the 5'end and the 3'end of the linear HBV are provided with complementary luciferase gene segments and specific sites recognized by the Phic31 recombinase, under the action of the Phic31 recombinase, cccDNA is formed through cyclization, and meanwhile, the complementary luciferase is also expressed; the luciferase provides a quick way for quickly detecting the active HBV cccDNA, and is suitable for high-throughput screening.
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Description

Technical field

[0001] The present invention relates to a system for inducing the formation of HBV cccDNA and a construction method. Specifically, a linear genome can produce HBV cccDNA containing luciferase in liver cancer cells expressing PhiC31 recombinase, and the luciferase provides rapid detection of active HBV cccDNA The shortcut is suitable for high-throughput screening. Background technique

[0002] Hepatitis B virus (hepatitis B virus, HBV) is a DNA virus belonging to the Hepadnaviridae family. The HBV genome is about 3.2 kb in length and encodes 7 functional proteins: HBV core antigen (HBcAg), e antigen (HBeAg), surface antigen (L, M, S HBsAg), polymerase (pol) and X protein (HBx). About 240 million people worldwide are infected with HBV, of which about 25 million are chronically infected (Global Hepatitis Report, 2017). Chronic infection will greatly increase the risk of liver cirrhosis and liver cancer, and more than 700,000 people die from HBV-related diseases ever...

Claims

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