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Identification and re-addressing of a transcriptionally permissive locus in the porcine genome

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Title Identification and re-addressing of a transcriptionally permissive locus in the porcine genome
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Creator Not Available
 
Subject Large animal model Active transgenesis Targeted integration Somatic cell nuclear transfer Livestock
 
Description Large animal model Active
transgenesis Targeted integration Somatic cell
nuclear transfer Livestock
Recently, we established the Sleeping
Beauty transposon system for germ line competent
transgenesis in the pig. Here, we extend this approach
to re-target a transposon-tagged locus for a sitespecific
gene knock-in, and generated a syngeneic
cohort of piglets carrying either the original transposon
or the re-targeted event. A Cre-loxP-mediated
cassette exchange of the tagging transposon with a
different reporter gene was performed, followed by
flow cytometric sorting and somatic cell nuclear
transfer of recombined cells. In parallel, the original
cells were employed in somatic cell nuclear transfer to
generate clone siblings, thereby resulting in a clone
cohort of piglets carrying different reporter transposons
at an identical chromosomal location. Importantly,
this strategy supersedes the need for an
antibiotic selection marker. This approach expands
the arsenal of genome engineering technologies in
domestic animals, and will facilitate the development
of large animal models for human diseases. Potentially,
the syngeneic cohort of pigs will be instrumental
for vital tracking of transplanted cells in preclinical
assessments of novel cell therapies.
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Date 2019-12-05T11:01:47Z
2019-12-05T11:01:47Z
2015-10-23
 
Type Article
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/28165
 
Language English
 
Relation Not Available;
 
Publisher Not Available