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Ungulates with genetically modified immune systems

A technology for ungulates and animals, applied in the field of ungulates with genetically modified immune systems, which can solve problems such as the unknown sequence and arrangement of immunoglobulin genes

Inactive Publication Date: 2009-03-18
REVIVICOR INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0045] Although some progress has been made in expressing human antibodies in cattle, there are still greater challenges in inactivating endogenous bovine Ig genes, increasing the expression level of human antibodies, and producing human antibody expression in other large animals, such as pigs, so The sequences and arrangements of the immunoglobulin genes of these other large animals are largely unknown

Method used

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  • Ungulates with genetically modified immune systems
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  • Ungulates with genetically modified immune systems

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0374] Example 1: Porcine heavy chain targeting and generation of porcine animals lacking heavy chain expression

[0375] A portion of the porcine Ig heavy chain locus was isolated from a 3X redundant porcine BAC library. Typically, a BAC library can be generated by fragmenting total porcine genomic DNA, which can then be used to obtain a BAC library representing at least three times the genome of the entire animal. BACs containing porcine heavy chain immunoglobulins are then selected by hybridization of probes selective for porcine heavy chain immunoglobulins described herein.

[0376]The sequence from the clone (Seq ID 1) was used to generate a primer complementary to part of the J region (this primer is represented by Seq ID No. 2). Separately, a primer complementary to a part of the mu constant region of the Ig heavy chain (this primer is represented by Seq ID No. 3) was designed. These primers were used to amplify a fragment of the porcine Ig heavy chain (represented by...

Embodiment 2

[0403] Example 2: Porcine kappa light chain targeting and generation of pigs lacking kappa light chain expression

[0404] A portion of the porcine Ig kappa chain locus was isolated from a 3X redundant porcine BAC library. Typically, a BAC library can be generated by fragmenting total porcine genomic DNA, which can then be used to obtain a BAC library representing at least three times the genome of the entire animal. BACs containing porcine kappa light chain immunoglobulins can then be selected by hybridization of probes selective for porcine kappa light chain immunoglobulins as described herein.

[0405] A fragment of porcine Ig light chain kappa was amplified using a primer complementary to part of the J region (this primer is represented by Seq ID No. 10) and a primer complementary to part of the kappa C region (this primer was represented by Seq ID No. 11). The resulting amplification primers were cloned into a plasmid vector and maintained in Stable2 cells at 30°C (Seq I...

Embodiment 3

[0425] The characterization of embodiment 3 porcine lambda loci

[0426] To disrupt or disable porcine lambda, targeting strategies have been devised that allow removal or disruption of regions of the lambda locus, including the concatemer of J to C expression cassettes. BAC clones containing portions of the porcine genome can be generated. A portion of the porcine Ig lambda chain locus was isolated from a 3X redundant porcine BAC library. Typically, the BAC library can be generated by fragmenting the total genomic DNA of the pig, which is then used to obtain a BAC library representing at least three times the genome of the intact animal. BACs containing porcine lambda chain immunoglobulin can then be selected by hybridization of probes selective for porcine lambda chain immunoglobulin as described herein.

[0427] BAC clones containing the λJ-C flanking regions (see Figure 3) can be independently fragmented and subcloned into plasmid vectors. Individual subclones have been...

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Abstract

The present invention provides ungulate animals, tissue and organs as well as cells and cell lines derived from such animals, tissue and organs, which lack expression of functional endogenous immunoglobulin loci. The present invention also provides ungulate animals, tissue and organs as well as cells and cell lines derived from such animals, tissue and organs, which express xenogenous, such as human, immunoglobulin loci. The present invention further provides ungulate, such as porcine genomic DNA sequence of porcine heavy and light chain immunogobulins. Such animals, tissues, organs and cells can be used in research and medical therapy. In addition, methods are provided to prepare such animals, organs, tissues, and cells.

Description

[0001] This application claims priority to US Provisional Application No. 60 / 621,433, filed October 22, 2004, which is incorporated herein by reference in its entirety. field of invention [0002] The invention provides ungulates, tissues and organs lacking expression of functional endogenous immunoglobulin loci, as well as cells and cell lines derived from such animals, tissues and organs. The invention also provides ungulates, tissues and organs expressing xenogenic (eg, human) immunoglobulin loci, and cells and cell lines derived from such animals, tissues and organs. The present invention also provides porcine genomic DNA sequences of heavy and light chain immunoglobulins of ungulates, such as porcine. Such animals, tissues, organs and cells can be used for research and medical treatment. In addition, methods of making such animals, organs, tissues and cells are provided. Background of the invention [0003] Antigens are agents or substances that are recognized by the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/00A01K67/033A01K67/027C12N15/00
CPCA01K2267/01C12N2800/204A01K2217/00A01K2227/108A01K2217/075A01K67/0276C12N15/8509C12N15/873A01K2227/102A01K2227/103A01K2227/101C12N2800/30C12N2800/206A01K2267/02A01K2267/025A01K2207/15Y02A50/30A01K67/027
Inventor K·威尔斯D·阿亚雷斯
Owner REVIVICOR INC
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