US 9,807,987 B2
Recombination activating gene 2 gene targeting vector, production of SCID-like miniature pigs by TALEN-mediated gene targeting and use thereof
Jin-Hoi Kim, Seoul (KR); Deug-Nam Kwon, Seoul (KR); Randall S. Prather, Columbia, MO (US); and Kiho Lee, Columbia, MO (US)
Assigned to KONKUK UNIVERSITY INDUSTRIAL COOPERATION CORP., Seoul (KR); and THE CURATORS OF THE UNIVERSITY OF MISSOURI, Columbia, MO (US)
Appl. No. 15/36,457
Filed by KONKUK UNIVERSITY INDUSTRIAL COOPERATION CORP., Seoul (KR); and THE CURATORS OF THE UNIVERSITY OF MISSOURI, Columbia, MO (US)
PCT Filed Nov. 13, 2014, PCT No. PCT/KR2014/010896
§ 371(c)(1), (2) Date May 13, 2016,
PCT Pub. No. WO2015/072760, PCT Pub. Date May 21, 2015.
Claims priority of application No. 10-2013-0137677 (KR), filed on Nov. 13, 2013.
Prior Publication US 2016/0366860 A1, Dec. 22, 2016
Int. Cl. A01K 67/027 (2006.01); C12N 15/877 (2010.01); C12N 15/90 (2006.01); C12N 5/074 (2010.01); C12N 15/85 (2006.01)
CPC A01K 67/0276 (2013.01) [A01K 67/0278 (2013.01); C12N 5/0696 (2013.01); C12N 15/8509 (2013.01); C12N 15/8778 (2013.01); C12N 15/907 (2013.01); A01K 2217/075 (2013.01); A01K 2227/108 (2013.01); A01K 2267/0387 (2013.01); C12N 2015/8536 (2013.01); C12N 2501/40 (2013.01); C12N 2501/602 (2013.01); C12N 2501/603 (2013.01); C12N 2501/604 (2013.01); C12N 2501/606 (2013.01); C12N 2501/608 (2013.01); C12N 2506/025 (2013.01); C12N 2510/00 (2013.01)] 10 Claims
 
1. A method for producing severe combined immunodeficiency (SCID)-like miniature pigs having a recombination activating gene (RAG) 2-allelic mutation, the method comprising:
inducing an allelic mutation in a pig RAG 2 gene by exposing the gene to a transcription activator-like effector nuclease (TALEN) that recognizes a TALEN-recognizing sequence region as set forth in SEQ ID NO. 1 in Chromosome 2 of the pig;
producing mutant embryos by using cells comprising the induced allelic mutation, wherein the producing is performed by somatic cell nuclear transplantation (SCNT); and
transferring the embryos to a surrogate gilt and allowing the embryos to gestate to term.