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Gene capturing carrier and method for establishment of full genome mutated embryonic stem cell bank

An embryonic stem cell and whole genome technology, applied in the biological field, can solve the problems of not being able to completely cover all genes, not being able to mutagenize genes, not being able to perform gene replacement, etc.

Inactive Publication Date: 2009-06-03
恒拓开源信息科技股份有限公司
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] One problem in conventional gene trapping technology is that gene replacement (such as replacing mouse gene with human gene) and introduction of gene point mutation cannot be carried out. Another outstanding defect is that it cannot mutagenize the gene necessary for embryonic development (can be lead to embryonic lethal), so that the established cell bank does not completely cover all the genes in the genome

Method used

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  • Gene capturing carrier and method for establishment of full genome mutated embryonic stem cell bank
  • Gene capturing carrier and method for establishment of full genome mutated embryonic stem cell bank
  • Gene capturing carrier and method for establishment of full genome mutated embryonic stem cell bank

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Embodiment Construction

[0027] (1) Construction of capture vector

[0028] The construction of the capture vector follows the steps of enzyme digestion, ligation, screening, and purification according to molecular biology techniques, and each element is cloned into the plasmid pBR322 in sequence, and the components are selected and sorted according to the capture vector required for construction. These elements include the neomycin resistance gene (neo) or β-galactosidase and neomycin resistance gene fusion resistance gene (βgeo); long terminal repeat (LTR); splice acceptor (SA); Splice donor (SD); poly(A) sequence; β-galactosidase (LacZ); phosphoglycerate kinase (PGK) promoter (PGKp); endosome entry sequence (IRES); FLP recombinase (fliprecombinase ) recognition site (frt-F3); Cre / lox recombination system recognition site (LoxP-Lox511), etc.

[0029] (2) Construction of a genome-wide mutant embryonic stem cell bank and analysis and positioning of the insertion point gene sequence

[0030] Embryoni...

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Abstract

The invention relates to a set of capturing carriers capable of full genome capturing and a method for establishment of a mutated embryonic stem cell bank using the set of capturing carriers, which belongs to the field of biotechnology. Four capturing carriers related in the invention can respectively capture normal expression genes, induces and captures unexpressed genes and embryo lethal genes in normal cell, so as to subject the genes of full genome to capturing and gene mutation and to establish the monogenically mutated embryonic stem cell bank that covers the full genome.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a set of capturing carriers, and a method for using the capturing carriers to carry out whole-genome gene capture and establish a whole-genome mutant embryonic stem cell bank. Background technique [0002] For three decades, advances in genetic technology have enabled extensive use of laboratory mice to study gene function in depth, enabling us to gain a deeper understanding of human health and disease. However, these developments in biological and genetic technology are still not fully utilized in biological and medical research to achieve breakthroughs in these fields. The key issue is to increase the scale and speed of mouse gene function research, and to develop a set of standards to have a standardized interpretation of these research results, and to identify those drug targets (genes) most relevant to medicine in a timely manner. A genetic technique that is readily available for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N15/867C12N5/10C40B50/06C40B40/08
Inventor 马越朱启超武洲钟伶桃李明
Owner 恒拓开源信息科技股份有限公司
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