Screening method of influenza virus related host gene mutants

An influenza virus and screening method technology, applied in the field of random mutation, can solve the problems of narrowing the scope of gene screening, lack of causal relationship between genes and influenza virus infection, omission of unknown genes and non-coding regions, etc.

Active Publication Date: 2021-03-26
THE THIRD MEDICAL CENT OF THE CHINESE PEOPLES LIBERATION ARMY GENERAL HOSPITAL
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

RNA interference, cDNA library, CRISPR/Cas9 mutation, and chip technology need to know the sequence of the target gene in advance, which will narrow the scope of gene screening, and often miss some unknown genes and non-coding regions; high-throughput sequencing and chip technology There is no clear causal relationship between the genes and influenza v

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Screening method of influenza virus related host gene mutants
  • Screening method of influenza virus related host gene mutants
  • Screening method of influenza virus related host gene mutants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1. Screening of influenza virus-related host gene mutants.

[0023] (1) Establishment of a random gene mutation library: Transfect human lung cancer cell lines with CAG-tTA plasmids to obtain Tet-off cell lines stably expressing transcriptional activators regulated by tetracycline; search vectors for genes and effectively express transposases Plasmids were co-transfected into Tet-off cell lines to obtain homozygous cell mutants for the whole genome; the specific method is as follows:

[0024] 1. Establish a human lung cancer A549 cell line stably expressing Tet-off.

[0025] 1) Installation of gene expression regulatory switch: digest, centrifuge, collect cells, resuspend and count cells, inoculate A549 cells into 6-well plates (3×10 5 / well) at 37°C, 5% CO 2 Cultivate overnight in the incubator; the next day, transfect 2 μg of the plasmid (CAG-tTA) containing tetracycline transcriptional activator (tTA) into A549 cells according to the instructions of FugeneH...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a screening method of influenza virus related host gene mutants, and belongs to the technical field of random mutation. In order to screen out host genes related to influenza viruses, the invention provides a screening method. The screening method comprises the following steps: 1) transfecting a human lung cancer cell strain with CAG-tTA plasmids to obtain a Tel-off cell strain of a transcription activator stably expressed and regulated by tetracycline; co-transfecting the Tel-off cell strain with a gene search vector and plasmids that effectively express transposase toobtain a whole-genome pulmonary epithelial cell gene mutation library; 2) infecting the mutation library obtained in the step 1) by using the H1N1-WSN influenza virus; and 3) extracting the genome DNA of the cell strain, resisting the H1N1-WSN influenza virus, obtained in the step 2), and carrying out sequencing by Splinkette PCR to obtain the influenza virus related host gene mutants. The screening method can be used for screening the influenza virus related host gene mutants.

Description

technical field [0001] The invention relates to a method for screening influenza virus-related host gene mutants, which belongs to the technical field of random mutations. Background technique [0002] Seasonal influenza is an acute respiratory infectious disease and a serious public health problem for all mankind. Its characteristics include easy outbreaks, easy spread, easy evolution, wide susceptible population, easy seasonal epidemics, and high incidence. Due to the high mutation rate of influenza virus and the recombination between viruses, it is difficult for the existing anti-influenza virus drugs and vaccines to effectively deal with the variable influenza virus. Compared with viral proteins, host factors involved in viral infection and expansion are less likely to mutate, and are important targets for intervening in viral replication and inhibiting host responses. Therefore, screening and identification of key host factors against influenza virus may be a breakthro...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/85C12N15/10C12Q1/70C12Q1/02C40B50/06C12R1/93
CPCC12N15/85C12N15/102G01N33/5005C40B50/06C12N2800/107C12N2800/90
Inventor 常德D·查尔斯S·罗克什
Owner THE THIRD MEDICAL CENT OF THE CHINESE PEOPLES LIBERATION ARMY GENERAL HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap