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

Humanized il-4 and il-4r alpha animals

An IL-4R and humanized technology, applied in the fields of instruments, peptides, animal husbandry, etc., can solve the problems of non-targeting and PD cannot be measured

Active Publication Date: 2017-03-01
REGENERON PHARM INC
View PDF8 Cites 18 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the PD of such therapeutic molecules cannot be properly determined in certain non-human animals because these therapeutic molecules do not target endogenous IL-4 or IL-4Rα proteins

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
  • Humanized il-4 and il-4r alpha animals
  • Humanized il-4 and il-4r alpha animals
  • Humanized il-4 and il-4r alpha animals

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0182] Replacement of endogenous mouse IL-4 gene by human IL-4 gene

[0183] 8.8 kb containing the coding portion of the human IL-4 gene from exon 1 to exon 4 (including the 3' untranslated region) starting from the ATG start codon and a part of the 3' region downstream of exon 4 The human IL-4 gene replacement spans 6.3 kb of the coding portion of exon 1 to exon 4 (including the 3' untranslated region) starting from the ATG start codon and a portion of the 3' region downstream of exon 4 Mouse IL-4 gene locus. see Figure 2A .

[0184] use Genetic engineering technology (see Valenzuela et al. (2003) High-throughput engineering of the mouse genome coupled with high-resolution expression analysis, Nature Biotech, 21(6):652-659), constructed in a single targeting step for the mouse Replacement with a targeting construct of the human IL-4 gene. Mouse and human IL-4 DNA were obtained from bacterial artificial chromosome (BAC) clones bMQ-41A12 and RP11-17K19, respectively. Br...

example 2

[0195] Replacement of endogenous mouse IL-4Rα ectodomain gene sequence by human IL-4Rα ectodomain gene sequence

[0196] A 15.6 kb human IL-4Rα gene replacement containing part of exon 1 starting at the ATG start codon to exon 5 and intron 5 of the human IL-4Rα gene spans the exon starting at the ATG start codon The 7.1 kb murine IL-4Rα gene locus from exon 1 to exon 5 and part of intron 5. Mouse exons 6 to 9 were retained; only exons 1 to 5 (ie, the extracellular domain) were humanized. see Figure 2B .

[0197] use Genetic engineering technology (see Valenzuela et al. (2003) High-throughput engineering of the mouse genome coupled with high-resolution expression analysis, Nature Biotech, 21(6):652-659), constructed in a single targeting step for the mouse Replacement with a targeting construct of the human IL-4Rα gene. Mouse and human IL-4Rα DNA were obtained from bacterial artificial chromosome (BAC) clones RP23-136G14 and RP11-166E24, respectively. Briefly, NotI linea...

example 3

[0208] Generation of double humanized IL-4 / IL-4Rα mice

[0209] Double humanized IL-4 / IL-4Ra (Il4 hu / hu / Il4ra hu / hu ) mice. ES cell clone MAID 803 including the humanized IL-4Rα gene and the neo box flanked by loxP sites was electroporated with a Cre expression vector to remove the neo box flanked by loxP sites to generate human ES cell clone MAID 1444 without the drug selection cassette (see Example 2) was derivatized with the IL-4Rα gene. The same targeting construct used to generate ES cell clone MAID 879 comprising the humanized IL-4 gene and a hygro cassette flanked by loxP sites (see Example 1) was electroporated into ES cell clone MAID 1444, to generate 879Het / 1444Het (Il4 + / hu / Il4ra + / hu ) ES cells, which were then electroporated with a Cre expression vector to remove the hygro box flanked by loxP sites to generate ES cell clones (MAID 1553 / 1444). The cassette-free ES cell clone MAID 1553 / 1444 was introduced into SW mouse embryos at the 8-cell stage to genera...

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

Non-human animals comprising a human or humanized IL-4 and / or IL-4R nucleic acid sequence are provided. Non-human animals that comprise a replacement of the endogenous IL-4 gene and / or IL-4R gene with a human IL-4 gene and / or IL-4R gene in whole or in part, and methods for making and using the non-human animals, are described. Non-human animals comprising a human or humanized IL-4 gene under control of non-human IL-4 regulatory elements is also provided, including non-human animals that have a replacement of non-human IL-4-encoding sequence with human IL-4-encoding sequence at an endogenous non-human IL-4 locus. Non-human animals comprising a human or humanized IL-4R gene under control of non-human IL-4R regulatory elements is also provided, including non-human animals that have a replacement of non-human IL-4R -encoding sequence with human or humanized IL-4R -encoding sequence at an endogenous non-human C IL-4R locus. Non-human animals comprising human or humanized IL-4 gene and / or IL-4R sequences, wherein the non-human animals are rodents, e.g., mice or rats, are provided.

Description

[0001] Cross References to Related Applications [0002] This patent application claims the benefit of priority to US Provisional Application No. 61 / 989,757, filed May 7, 2014, the entire contents of which are incorporated herein by reference. [0003] SEQUENCE LISTINGS INCORPORATE BY REFERENCE [0004] The sequence listing as an ASCII text file, 16 KB named 31260_SEQ.txt, created on May 7, 2015, and submitted to the US Patent and Trademark Office via EFS-Web is incorporated herein by reference. technical field [0005] Disclosed herein are non-human animals comprising nucleic acid sequences encoding IL-4 and / or IL-4Rα proteins comprising human sequences. Also disclosed herein are transgenic non-human animals comprising a wholly or partially human IL-4 and / or IL-4Ra gene. The invention also discloses non-human animals expressing human or humanized IL-4 and / or IL-4Rα protein. In addition, the present invention discloses methods for making and using non-human animals comprisi...

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): A01K67/027
CPCA01K67/0278C07K14/5406C07K14/7155A01K2267/0368A01K2227/105A01K2217/072A01K2217/20A01K2267/01A01K2267/0393A01K2207/15G01N33/5088C12N15/8509
Inventor L-H·王Y·薛A·J·墨菲S·史蒂文斯
Owner REGENERON PHARM INC
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