PMID- 25437445
OWN - NLM
STAT- MEDLINE
DCOM- 20150915
LR  - 20181113
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 9
IP  - 12
DP  - 2014
TI  - Generation of recombination activating gene-1-deficient neonatal piglets: a model 
      of T and B cell deficient severe combined immune deficiency.
PG  - e113833
LID - 10.1371/journal.pone.0113833 [doi]
LID - e113833
AB  - Although severe combined immune deficiency (SCID) is a very important research 
      model for mice and SCID mice are widely used, there are only few reports 
      describing the SCID pig models. Therefore, additional research in this area is 
      needed. In this study, we describe the generation of Recombination activating 
      gene-1 (Rag-1)-deficient neonatal piglets in Duroc breed using somatic cell 
      nuclear transfer (SCNT) with gene targeting and analysis using 
      fluorescence-activated cell sorting (FACS) and histology. We constructed porcine 
      Rag-1 gene targeting vectors for the Exon 2 region and obtained 
      heterozygous/homozygous Rag-1 knockout cell colonies using SCNT. We generated two 
      Rag-1-deficient neonatal piglets and compared them with wild-type neonatal 
      piglets. FACS analysis showed that Rag-1 disruption causes a lack of 
      Immunoglobulin M-positive B cells and CD3-positive T cells in peripheral blood 
      mononuclear cells. Consistent with FACS analysis, histological analysis revealed 
      structural defects and an absence of mature lymphocytes in the spleen, mesenteric 
      lymph node (MLNs), and thymus in Rag-1-deficient piglets. These results confirm 
      that Rag-1 is necessary for the generation of lymphocytes in pigs, and 
      Rag-1-deficient piglets exhibit a T and B cell deficient SCID (T-B-SCID) 
      phenotype similar to that of rodents and humans. The T-B-SCID pigs with Rag-1 
      deficiency generated in this study could be a suitably versatile model for 
      laboratory, translational, and biomedical research, including the development of 
      a humanized model and assessment of pluripotent stem cells.
FAU - Ito, Tetsuya
AU  - Ito T
AD  - Pig Breeding Laboratory, Central Research Institute for Feed and Livestock, 
      ZEN-NOH (National Federation of Agricultural Co-operative Associations), 
      Hokkaido, Japan.
FAU - Sendai, Yutaka
AU  - Sendai Y
AD  - Biological Sciences Section, Central Research Institute for Feed and Livestock, 
      ZEN-NOH (National Federation of Agricultural Co-operative Associations), Ibaraki, 
      Japan.
FAU - Yamazaki, Satoshi
AU  - Yamazaki S
AD  - Division of Stem Cell Therapy, The Institute of Medical Science, The University 
      of Tokyo, Tokyo, Japan; Japan Science and Technology Agency, Exploratory Research 
      for Advanced Technology (ERATO), Nakauchi Stem Cell and Organ Regeneration 
      Project, Tokyo, Japan.
FAU - Seki-Soma, Marie
AU  - Seki-Soma M
AD  - Zen-noh Institute of Animal Health, ZEN-NOH (National Federation of Agricultural 
      Co-operative Associations), Chiba, Japan.
FAU - Hirose, Kensuke
AU  - Hirose K
AD  - Pig Breeding Laboratory, Central Research Institute for Feed and Livestock, 
      ZEN-NOH (National Federation of Agricultural Co-operative Associations), 
      Hokkaido, Japan.
FAU - Watanabe, Motoo
AU  - Watanabe M
AD  - Division of Stem Cell Therapy, The Institute of Medical Science, The University 
      of Tokyo, Tokyo, Japan; Japan Science and Technology Agency, Exploratory Research 
      for Advanced Technology (ERATO), Nakauchi Stem Cell and Organ Regeneration 
      Project, Tokyo, Japan.
FAU - Fukawa, Kazuo
AU  - Fukawa K
AD  - Pig Breeding Laboratory, Central Research Institute for Feed and Livestock, 
      ZEN-NOH (National Federation of Agricultural Co-operative Associations), 
      Hokkaido, Japan.
FAU - Nakauchi, Hiromitsu
AU  - Nakauchi H
AD  - Division of Stem Cell Therapy, The Institute of Medical Science, The University 
      of Tokyo, Tokyo, Japan; Japan Science and Technology Agency, Exploratory Research 
      for Advanced Technology (ERATO), Nakauchi Stem Cell and Organ Regeneration 
      Project, Tokyo, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20141201
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Homeodomain Proteins)
RN  - 128559-51-3 (RAG-1 protein)
SB  - IM
MH  - Animals
MH  - Animals, Newborn
MH  - B-Lymphocytes/*metabolism
MH  - Cells, Cultured
MH  - Disease Models, Animal
MH  - Fibroblasts/cytology
MH  - Gene Knockout Techniques
MH  - Homeodomain Proteins/*genetics
MH  - Nuclear Transfer Techniques
MH  - Severe Combined Immunodeficiency/*genetics/immunology/pathology
MH  - Swine
MH  - T-Lymphocytes/*metabolism
PMC - PMC4249935
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2014/12/02 06:00
MHDA- 2015/09/16 06:00
PMCR- 2014/12/01
CRDT- 2014/12/02 06:00
PHST- 2014/07/28 00:00 [received]
PHST- 2014/10/30 00:00 [accepted]
PHST- 2014/12/02 06:00 [entrez]
PHST- 2014/12/02 06:00 [pubmed]
PHST- 2015/09/16 06:00 [medline]
PHST- 2014/12/01 00:00 [pmc-release]
AID - PONE-D-14-33789 [pii]
AID - 10.1371/journal.pone.0113833 [doi]
PST - epublish
SO  - PLoS One. 2014 Dec 1;9(12):e113833. doi: 10.1371/journal.pone.0113833. 
      eCollection 2014.