Format

Send to

Choose Destination
Reprod Biol Endocrinol. 2016 Nov 3;14(1):74.

Biallelic modification of IL2RG leads to severe combined immunodeficiency in pigs.

Author information

1
MGENPLUS Biotechnology Research Institute, Seoul, 08511, South Korea.
2
Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg, VA, USA.
3
Department of Large Animal Clinical Sciences, Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, VA, USA.
4
Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg, VA, USA. kiholee@exchange.vt.edu.
5
MGENPLUS Biotechnology Research Institute, Seoul, 08511, South Korea. parkmgen@hanmail.net.
6
Department of Animal Science & Technology, Sunchon National University, Suncheon, 57922, South Korea. parkmgen@hanmail.net.

Abstract

BACKGROUND:

Pigs with SCID can be a useful model in regenerative medicine, xenotransplantation, and cancer cell transplantation studies. Utilizing genome editing technologies such as CRISPR/Cas9 system allows us to generate genetically engineered pigs at a higher efficiency. In this study, we report generation and phenotypic characterization of IL2RG knockout female pigs produced through combination of CRISPR/Cas9 system and SCNT. As expected, pigs lacking IL2RG presented SCID phenotype.

METHODS:

First, specific CRISPR/Cas9 systems targeting IL2RG were introduced into developing pig embryos then the embryos were transferred into surrogates. A total of six fetuses were obtained from the embryo transfer and fetal fibroblast cell lines were established. Then IL2RG knockout female cells carrying biallelic genetic modification were used as donor cells for SCNT, followed by embryo transfer.

RESULTS:

Three live cloned female piglets carrying biallelic mutations in IL2RG were produced. All cloned piglets completely lacked thymus and they had a significantly reduced level of mature T, B and NK cells in their blood and spleen.

CONCLUSIONS:

Here, we generated IL2RG knockout female pigs showing phenotypic characterization of SCID. This IL2RG knockout female pigs will be a promising model for biomedical and translational research.

KEYWORDS:

CRISPR/Cas9; IL2RG; Knockout pigs; Severe Combined Immunodeficiency (SCID); Somatic cell nuclear transfer (SCNT)

PMID:
27809915
PMCID:
PMC5095964
DOI:
10.1186/s12958-016-0206-5
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for BioMed Central Icon for PubMed Central
Loading ...
Support Center