Targeted Gene Editing in Porcine Spermatogonia.
CRISPR/Cas9
gene targeting
homology directed repair
homology-mediated end joining
pig
spermatogonia
Journal
Frontiers in genetics
ISSN: 1664-8021
Titre abrégé: Front Genet
Pays: Switzerland
ID NLM: 101560621
Informations de publication
Date de publication:
2020
2020
Historique:
received:
09
11
2020
accepted:
31
12
2020
entrez:
15
2
2021
pubmed:
16
2
2021
medline:
16
2
2021
Statut:
epublish
Résumé
To study the pathophysiology of human diseases, develop innovative treatments, and refine approaches for regenerative medicine require appropriate preclinical models. Pigs share physiologic and anatomic characteristics with humans and are genetically more similar to humans than are mice. Genetically modified pigs are essential where rodent models do not mimic the human disease phenotype. The male germline stem cell or spermatogonial stem cell (SSC) is unique; it is the only cell type in an adult male that divides and contributes genes to future generations, making it an ideal target for genetic modification. Here we report that CRISPR/Cas9 ribonucleoprotein (RNP)-mediated gene editing in porcine spermatogonia that include SSCs is significantly more efficient than previously reported editing with TALENs and allows precise gene editing by homology directed repair (HDR). We also established homology-mediated end joining (HMEJ) as a second approach to targeted gene editing to enable introduction of larger transgenes and/or humanizing parts of the pig genome for disease modeling or regenerative medicine. In summary, the approaches established in the current study result in efficient targeted genome editing in porcine germ cells for precise replication of human disease alleles.
Identifiants
pubmed: 33584819
doi: 10.3389/fgene.2020.627673
pmc: PMC7876475
doi:
Types de publication
Journal Article
Langues
eng
Pagination
627673Subventions
Organisme : NICHD NIH HHS
ID : R01 HD091068
Pays : United States
Organisme : NIH HHS
ID : R01 OD016575
Pays : United States
Organisme : NIGMS NIH HHS
ID : R43 GM108150
Pays : United States
Organisme : NIGMS NIH HHS
ID : R44 GM108150
Pays : United States
Informations de copyright
Copyright © 2021 Webster, Bondareva, Solin, Goldsmith, Su, Lara, Carlson and Dobrinski.
Déclaration de conflit d'intérêts
DW, SS, TG, DC, and ID are employees and/or shareholders in Recombinetics, Inc., a company that commercializes gene editing in livestock. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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