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

Two step cluster deletion and humanisation

A humanized and mouse technology, applied in the field of humanized mice, to achieve the effect of promoting production and reducing the risk of cumulative damage

Inactive Publication Date: 2010-10-06
ITI SCOTLAND
View PDF6 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no indication that this technique can be used to humanize non-segmented genes, where the presence of nucleic acid sequences encoding remaining site-specific recombination sites in the gene should abrogate its transcriptional capacity

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
  • Two step cluster deletion and humanisation
  • Two step cluster deletion and humanisation
  • Two step cluster deletion and humanisation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0121] Example 1: Humanization of CYP3A4 using the corresponding human promoter and mouse CYP3A cluster knockout

[0122] The mouse CYP3A cluster is flanked by loxP and FRT sites, and the CYP3A4 expression cassette is inserted into one end of the mouse cluster, as in Figure 7 , thus allowing the expression of human CYP3A4 under the control of the 13 kb human CYP3A4 promoter.

[0123] The loxP sequence element included in the targeting vector allows Cre-mediated deletion of the mouse CYP3A cluster, thereby allowing expression of CYP3A4 in the absence of the corresponding mouse gene (see Figure 7 ).

[0124] Subsequently, the FRT sequence elements allow Flp-mediated deletion of the human expression cassette, resulting in a complete knockout at the mouse CYP3A locus ( Figure 7 ).

Embodiment 2

[0125] Example 2: Humanization of CYP2C9 using the corresponding human promoter and mouse CYP2C cluster knockout

[0126] The mouse CYP2C cluster is flanked by loxP and FRT sites, and a CYP2C9 expression cassette is inserted into one end of the mouse cluster, as in Figure 8 , thus allowing the expression of human CYP2C9 under the control of the 12 kb human CYP2C9 promoter.

[0127] The loxP sequence element included in the targeting vector allows Cre-mediated deletion of the mouse CYP2C cluster, thereby allowing expression of CYP2C9 in the absence of the corresponding mouse gene (see Figure 8 ).

[0128] Subsequently, the FRT sequence elements allow Flp-mediated deletion of the human expression cassette, resulting in a complete knockout at the mouse CYP2C locus ( Figure 8 ).

Embodiment 3

[0129] Example 3: Humanization of CYP2D6 using the corresponding human promoter and mouse CYP2D cluster knockout

[0130] The mouse CYP2D cluster is flanked by loxP and FRT sites, and the CYP2D6 expression cassette is inserted into one end of the mouse cluster, as in Figure 9 , thus allowing the expression of human CYP2D6 under the control of the 9 kb human CYP2D6 promoter.

[0131] The loxP sequence element included in the targeting vector allows Cre-mediated deletion of the mouse Cyp2d cluster, thereby allowing expression of CYP2D6 in the absence of the corresponding mouse gene. Subsequently, the FRT sequence elements allow Flp-mediated deletion of the human expression cassette, resulting in a complete knockout at the mouse CYP2D locus.

[0132] The use of specific primers allows analysis of mouse chromosomes by PCR. The results demonstrate successful introduction of the desired human replacement CYP2D gene sequence and deletion of the mouse Cyp2d cluster (see Figure 10...

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

This invention relates to a method for humanising a mouse. In particular, the invention relates to a method for replacing a cluster of mouse genes with single or multiple genes from the corresponding human cluster using a combination of homologous recombination and site-specific recombination.

Description

field of invention [0001] The present invention relates, generally, to methods for humanizing mice. In particular, the invention relates to methods for replacing a mouse gene cluster with one or more genes from a corresponding human cluster using a combination of homologous and site-specific recombination. Background of the invention [0002] Mouse models are invaluable tools for studying human disease and are widely used to study the progression of many diseases, and to test potential treatments. Conventionally, transgenic mice have been generated by pronuclear injection of exogenous DNA. More recently, mice have been generated by fusing embryonic stem cells with cells containing bacterial artificial chromosomes (BACs) or yeast artificial chromosomes (YACs) containing exogenous genes of interest and Selectable markers for assessing the integration of exogenous DNA segments into the genome of blast cells are described, for example, in WO94 / 02602. This method relies on the...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/90A01K67/027C12N9/02
CPCA01K2217/075C12N9/0071A01K2227/105A01K2217/05A01K2217/072A01K2267/03A01K67/0275C12N15/8509A01K2217/00A01K2207/15
Inventor 尼科·舍尔
Owner ITI SCOTLAND
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