Apancreatic pigs cloned using Pdx1-disrupted fibroblasts created via TALEN-mediated mutagenesis
dc.contributor.author
Kang, Jin-Dan
dc.contributor.author
Kim, Hyojin
dc.contributor.author
Jin, Long
dc.contributor.author
Guo, Qing
dc.contributor.author
Cui, Cheng-Du
dc.contributor.author
Li, Wen-Xue
dc.contributor.author
Kim, Seokjoong
dc.contributor.author
Kim, Jin-Soo
dc.contributor.author
Yin, Xi-Jun
dc.date.accessioned
2018-02-23T14:37:59Z
dc.date.available
2018-02-21T03:00:33Z
dc.date.available
2018-02-23T14:37:59Z
dc.date.issued
2017
dc.identifier.issn
1949-2553
dc.identifier.other
10.18632/oncotarget.23301
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/242638
dc.identifier.doi
10.3929/ethz-b-000242638
dc.description.abstract
Pancreatic and duodenal homeobox 1 (PDX1) plays a crucial role in pancreas development, β-cell differentiation, and maintenance of mature β-cell function. In this study, we designed a strategy to produce PDX1-knockout (KO) pigs. A transcription activator-like effector nuclease (TALEN) pair targeting exon 1 of the swine PDX1 gene was constructed. Porcine fetal fibroblasts (PFFs) were transfected with the TALEN plasmids plus a surrogate reporter plasmid. PDX1-mutated PFFs were enriched by magnetic separation and used to produce homozygous PDX1-KO pigs via a two-step somatic cell nuclear transfer (SCNT) cloning process. In the first SCNT step, we obtained eight fetuses, established PFF cell lines, and analyzed PDX1 gene mutations by T7 endonuclease 1 assays and Sanger sequencing. Five fetuses showed mutations at the PDX1 loci with two biallelic mutations and three monoallelic mutations (mutation rate of 62.5%). In the second step, a PDX1 biallelic mutant PFF cell line with a 2 bp deletion in one allele and a 4 bp insertion in the other allele was used as a donor to generate cloned pigs via SCNT. From 462 cloned embryos transferred into two surrogates, nine live piglets were delivered. These piglets at birth were not clearly distinguishable phenotypically from wild-type piglets, but soon developed severe diarrhea and vomiting and all died within 2 days after birth. Dissection of PDX1-KO piglets revealed that the liver, gallbladder, spleen, stomach, common bile duct, and other viscera were present and normal, but the pancreas was absent in all cases.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Impact Journals
dc.rights.uri
http://creativecommons.org/licenses/by/3.0/
dc.subject
PDX1
en_US
dc.subject
TALEN
en_US
dc.subject
SCNT
en_US
dc.subject
pancreas
en_US
dc.subject
pig
en_US
dc.title
Apancreatic pigs cloned using Pdx1-disrupted fibroblasts created via TALEN-mediated mutagenesis
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 3.0 Unported
ethz.journal.title
Oncotarget
ethz.journal.volume
8
en_US
ethz.journal.issue
70
en_US
ethz.pages.start
115480
en_US
ethz.pages.end
115489
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.publication.place
Albany, NY
ethz.publication.status
published
en_US
ethz.date.deposited
2018-02-21T03:00:38Z
ethz.source
WOS
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2018-02-23T14:38:35Z
ethz.rosetta.lastUpdated
2024-02-02T04:00:43Z
ethz.rosetta.versionExported
true
ethz.COinS
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Journal Article [130185]