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Pathogen-resistant animals having modified aminopeptidase n (ANPEP) genes

A gene and animal technology, applied in a field that further involves the production of anti-pathogen non-human animals or non-human animal lineages, can solve the problems of poor understanding of pathogenesis and no reliable technology

Inactive Publication Date: 2021-02-02
UNIV OF MISSOURI BOARD OF CURATORS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The difficulty and much of the expense associated with this strategy is that the pathogenesis of persistent PRRSV infection is poorly understood and therefore there are no reliable techniques for identifying persistently infected pigs.

Method used

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  • Pathogen-resistant animals having modified aminopeptidase n (ANPEP) genes
  • Pathogen-resistant animals having modified aminopeptidase n (ANPEP) genes
  • Pathogen-resistant animals having modified aminopeptidase n (ANPEP) genes

Examples

Experimental program
Comparison scheme
Effect test

example

[0561]The following non-limiting examples are provided to further illustrate the invention.

example 1

[0563]Example 1: Using CRISPR / Cas9 system to produce genetically engineered pigs from oocytes and embryos sourced in vitro

[0564]Recent reports describe homing endonucleases, such as zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN) and clustered regularly spaced short palindrome repeats (CRISPR) / CRISPR associations (Cas9) components in the system, the report suggests that genetic engineering (GE) of pigs may now be more efficient. Targeted homing endonuclease can induce double-strand breaks (DSB) at specific locations in the genome, and if donor DNA is provided, it can be achieved by non-homologous end joining (NHEJ) or stimulation of homologous recombination (HR) Cause random mutations. If the donor DNA is provided along with the targeted nuclease, homing endonuclease can be used to achieve targeted modification of the genome through HR. After introducing specific modifications into somatic cells, these cells are used to produce GE pigs for various ...

example 2

[0663]Example 2: Pigs with modified chromosomal sequence in the gene encoding CD163 protein have increased resistance to PRRSVplus

[0664]Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) ravaged the pig industry in the past quarter of a century. In this example, it is shown that the CD163-null animals did not show all the clinical signs of infection, lung pathology, viremia, or antibody production that are characteristic of PRRSV infection. It has not only confirmed that PRRSV enters the intermediary; but if animals produced in a similar manner are allowed to enter the food supply, strategies to prevent major economic losses and animal suffering have been described.

[0665]Materials and methods

[0666]Genotyping

[0667]Genotyping is based on both DNA sequencing and mRNA sequencing. The genotype of this male animal has an 11 bp deletion in one allele, and 45 amino acids of domain 5 are predicted when translated, resulting in an early stop codon at amino acid 64. In another allele,...

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Abstract

Livestock animals and offspring thereof comprising at least one modified chromosomal sequence in a gene encoding an aminopeptidase N (ANPEP) protein are provided. Animal cells that contain such modified chromosomal sequences are also provided. The animals, offspring, and cells have increased resistance to pathogens, including transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV). The animals, offspring, and cells can optionally further comprise at least one modified chromosomal sequence in a gene encoding a CD163 protein and / or a SIGLEC1 protein. Methods for producing pathogen-resistant non-human animals or lineages of non-human animals are also provided.

Description

[0001]Electronically submitted sequence list reference[0002]The official copy of the sequence list-that is, the ASCII format sequence list created on April 26, 2019 and the file size is 318.7 kilobytes named "(16UMC002-WO) sequence list archived as 4.26.19"- Submit electronically through the EFS network and submit it at the same time as this manual. The sequence list contained in this ASCII format document is a part of this specification and is fully incorporated herein by reference.Technical field[0003]The present invention relates to livestock animals and their descendants that contain at least one modified chromosomal sequence in the gene encoding aminopeptidase N (ANPEP) protein. The present invention further relates to an animal cell containing at least one modified chromosomal sequence in the gene encoding the ANPEP protein. Animals and cells have increased resistance to pathogens, including transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV)....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/48A61K39/00
CPCA01K67/0275C12N2310/20A01K2217/072A01K2227/108A01K2267/02C12N15/90C12N9/485C12Y304/11C07K14/70596
Inventor R·S·普莱特尔K·D·韦尔斯K·M·惠特沃思
Owner UNIV OF MISSOURI BOARD OF CURATORS
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