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

Human artificial chromosome containing human antibody lambda light chain

A light chain gene and human antibody technology, applied in genetic engineering, antibodies, anti-animal/human immunoglobulin, etc., can solve the problem of low chimerism in chimeric mice

Inactive Publication Date: 2006-06-21
KYOWA HAKKO KIRIN CO LTD +1
View PDF6 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In contrast, when a fragment of human chromosome 22 was introduced, chimeric mice obtained in most cases had a low degree of chimerism (i.e., 50% or less)

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
  • Human artificial chromosome containing human antibody lambda light chain
  • Human artificial chromosome containing human antibody lambda light chain
  • Human artificial chromosome containing human antibody lambda light chain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] [Example 1] Production of expression cassette vector pTELhisD

[0078] The expression cassette vector pTELPuro (Kuroiwa et al., Nature Biotech., 18: 1086-, 2000) was cut with restriction enzyme NotI (Boehringer), and blunted at 72°C for 5 minutes using a DNA blunting kit (Toyobo Co., Ltd.). After blunting, it was dephosphorylated at 65° C. for 1 hour using bacterial-derived alkaline phosphatase (Takara Shuzo Co., Ltd.). Thereafter, a restriction enzyme BglII linker (Takara Shuzo Co., Ltd.) was added, and ligation was performed using a ligation kit (Takara Shuzo Co., Ltd.). The plasmid pTELBg in which the BgIII linker replaces the PGKPuro expression cassette in the pTELPuro plasmid was thus generated. This plasmid was cut with restriction enzyme BglII, and dephosphorylated in the same manner. Thereafter, it was purified by gel filtration using CHROMA SPIN-TE 400 (Clontech). Subsequently, the hisD fragment was excised from plasmid #1-132 (provided by Professor Shun-ich...

Embodiment 2

[0079] [Example 2] Production of targeting vector pTELhisDλI

[0080] The targeting vector pTELhisDλI for inserting the human telomere sequence into the AP000344 region very close to the Igλ locus on human chromosome 22 and on the telomere side (approximately 400 Kb telomere side) was produced in the following manner. Initially, the AP000344 genomic region was amplified by PCR using the following primers.

[0081] 1269D1-F; 5'-TCGAGGATCCGACAAGTTCTCTTCTCTTTTCCTTCTGCCC-3' (SEQ ID NO: 1)

[0082] 1269D1-R; 5'-TCGAGGATCCGCTGCTAAGCTACTGTTCTCTTTTTTCCCC-3' (SEQ ID NO: 2)

[0083] PCR was performed using GeneAmp 9600 (manufactured by Perkin-Elmer) as a thermal cycler, LA Taq (Takara Shuzo Co., Ltd.) as Taq polymerase, and ligation buffer and dNTP (dATP, dCTP, dGTP, dTTP). For temperature and cycle conditions, after heat denaturation at 94°C for 1 minute, 35 cycles were performed at 98°C for 10 seconds and 68°C for 11 minutes. The PCR product was treated with proteinase K (Gibco), ...

Embodiment 3

[0084] [Example 3] Production of targeting vector p5531oxPHyg

[0085] The targeting vector p553loxPHyg, which is used to insert the Cre recombinase recognition sequence loxP sequence into AP000553 very close to the Igλ locus on human chromosome 22 and on the centromere side (about 300Kb centromere side), was produced in the following manner Area. Initially, the AP000553 genomic region was amplified by PCR using the following primers.

[0086] 553-F3; 5'-TCGAGTCGACTGTAGCTGACTTTAGCCACCCACAAGTAC-3' (SEQ ID NO: 3)

[0087] 553-R3; 5'-TCGAGTCGACCTTGCTGATTATACCTCATCCTCCTTCCCTC-3' (SEQ ID NO: 4)

[0088] PCR was performed using GeneAmp 9600 (manufactured by Perkin-Elmer) as a thermal cycler, LA Taq (Takara Shuzo Co., Ltd.) as Taq polymerase, and ligation buffer and dNTP (dATP, dCTP, dGTP, dTTP). For temperature and cycle conditions, after heat denaturation at 94°C for 1 minute, 35 cycles were performed at 98°C for 10 seconds and 68°C for 15 minutes. The PCR product was treated ...

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 present invention relates to a human artificial chromosome which is genetically transmissible to the next generation with high efficiency and the method for using the same. More specifically, the present invention relates to: a human artificial chromosome in which an about 3.5 Mb to about 1 Mb region containing an antibody » light chain gene derived from human chromosome 22 is bound to a chromosome fragment which is transmissible to a progeny through a germ line of a non-human animal, said chromosome fragment is derived from another human chromosome; a non-human animal carrying the human artificial chromosome and an offspring thereof; a method for producing the non-human animal; a method for producing a human antibody using the non-human animal or an offspring thereof; and a human antibody-producing mouse carrying the human artificial chromosome.

Description

[0001] This application is a divisional application, the filing date of the original application is May 10, 2002, the application number is 02813520.2, and the title of the invention is "Human Artificial Chromosome Containing Human Antibody λ Light Chain Gene". technical field [0002] The present invention relates to a human artificial chromosome that can be efficiently genetically transmitted to the next generation by modifying the chromosome or a fragment thereof, a non-human animal that can efficiently genetically transmit the human artificial chromosome to the next generation and its offspring, the non-human animal or A method for producing antibodies by its progeny and a mouse for producing human antibodies. technical background [0003] A technique for producing chimeric animals from hybrid cells obtained by fusion of minicells containing chromosome fragments and pluripotent cells has been developed (WO 97 / 07671). The technique enables the production of non-human anim...

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): A01K67/027C12N15/09A61K48/00C07K16/00C07K16/24C12N5/12C12N15/02C12N15/13C12N15/85C12N15/87C12N15/90C12P21/00C12P21/08
CPCA01K67/0275A01K2207/15A01K2217/00A01K2217/05A01K2227/105A01K2267/01A01K2267/0381A61K48/00A61K2039/505C07K16/00C07K16/243C07K2317/21C12N15/87C12N15/90C12N2800/30A01K67/0278C12N15/8509C12N15/09C07K16/18C12N5/16A01K67/0276A01K2217/052A01K2217/054A01K2217/15
Inventor 黑岩义巳富塚一磨吉田均石田功
Owner KYOWA HAKKO KIRIN CO LTD
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