Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

TALEN-mediated vector for knocking out goat BLG through gene targeting and recombinant cell

A technology of gene targeting and gene knockout, which is applied in the field of animal genetic engineering, can solve problems such as the impact on the safety of individual organisms and complex workload, and achieve the effect of improving the efficiency of gene targeting

Active Publication Date: 2015-06-24
NORTHWEST A & F UNIV +1
View PDF3 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Screening for loss-of-function mutants is complicated by the random and unpredictable nature of modifications obtained by this method
In addition, the random integration of helper vectors has a great impact on the safety of individual organisms

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
  • TALEN-mediated vector for knocking out goat BLG through gene targeting and recombinant cell
  • TALEN-mediated vector for knocking out goat BLG through gene targeting and recombinant cell
  • TALEN-mediated vector for knocking out goat BLG through gene targeting and recombinant cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0038] In order to improve the gene targeting efficiency of knocking out goat β-lactoglobulin gene, the present invention uses TALEN (transcription activator-like effector nuclease) to identify the first goat β-lactoglobulin (C.hircus beta-lactoglobulin) gene A site on the exon region generates a double-strand break after cutting, and initiates DNA double-strand break repair. At the same time, a homology arm vector is introduced, and the DNA recombination repair process triggered by DNA double-strand breaks is used to make the assembled target DNA sequence The goat β-lactoglobulin gene is specifically inserted to realize the modification and knockout of the goat β-lactoglobulin gene. The following are the construction of TALEN eukaryotic expression vector, TALEN expression vector as template for in vitro transcription of mRNAs, construction of BLG gene knockout vector with shorter homology arms, co-transfection of goat fetal fibroblasts, drug screening and identification A fur...

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 TALEN-mediated vector for knocking out goat BLG through gene targeting and a recombinant cell. A TALEN eukaryotic expression vector aiming at a first exon of a goat beta-lactoglobulin gene and a BLG gene knockout vector containing short homologous arms are designed and constructed according to the goat beta-lactoglobulin gene and are jointly transfect goat fetal fibroblast together with mRNAs obtained by in vitro transcription based on TALEN expression vector as a template, and a BLG gene knockout cell is obtained after drug screening and identification. According to the invention, the gene targeting efficiency is greatly improved by utilizing a TALEN technology; meanwhile, the utilization of the short homologous arms facilitates more convenient and more efficient screening of the targeted cells; random integration of the TALEN expression vector in genomes is avoided by utilizing mRNAs.

Description

technical field [0001] The invention belongs to the technical field of animal genetic engineering, and relates to a vector and a recombinant cell for knocking out goat BLG based on TALEN-mediated gene targeting. Background technique [0002] β-lactoglobulin (BLG) is the main whey protein in ruminant milk, about 56%-60%, basically absent in human and rodent milk (Kontopidis G et al.2004), is One of the main allergens causing milk allergy in infants and young children. At present, there are many biochemical methods to reduce the allergenicity of BLG, such as heat treatment, enzymatic hydrolysis, glycosylation, and fermentation. Although these methods have a certain effect on reducing BLG allergy, they cannot fundamentally solve the sensitization problem of BLG. With the development of molecular biology technology, RNA interference technology can effectively reduce BLG in milk, and its interference efficiency can reach 100%. However, the use of RANi to produce transgenic ani...

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): C12N15/85C12N15/55C12N15/12C12N5/10C12N15/873
Inventor 张涌靳亚平崔趁趁刘军葛恒涛宋玉洁
Owner NORTHWEST A & F UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com