TALEN‐mediated gene targeting in porcine spermatogonia

TALEN‐mediated gene targeting in porcine spermatogonia AbbreviationsCRISPR/Cas9Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR‐associated‐9DMDDuchenne Muscular DystrophyDMEMDulbecco's Modified Eagle MediumDPBSDulbecco's PhoSphate Buffered SalineEDTAethylenediaminetetra acetic acidESCembryonic stem cellFACSfluorescence activated cell sortingGFPgreen fluorescent proteinHRhomologous recombinationiPSCinduced pluripotent stem cellMACSmagnetic activated cell sortingNHEJnon‐homologous end joiningSCNTsomatic cell nuclear transferSSCspermatogonial stem cellTALENTranscription Activator‐like Effector NucleaseUCH‐L1ubiquitin carboxyterminal hydrolase L‐1ZFNzinc finger nucleaseINTRODUCTIONSpermatogonia represent a diploid germ cell population in the seminiferous tubules which undergo mitotic division during spermatogenesis to give rise to primary spermatocytes. Spermatogonial stem cells (SSCs) are a subset of undifferentiated type A spermatogonia which have the capacity to self‐renew to maintain the SSC pool as well as to differentiate to ultimately form sperm. SSCs represent a rare cell population, constituting ∼0.03% of all germ cells in the adult mouse testis (Tegelenbosch & de Rooij, ). The lack of definite phenotypical, morphological, or biochemical markers that can unequivocally identify SSCs makes it impossible to isolate SSCs for in vitro studies. However, SSCs can be relatively enriched from a testicular cell suspension by various in vivo and/or in vitro approaches.For in vivo enrichment, neonatal, and prepubertal testes serve as the preferred source for harvesting germ cells as gonocytes/spermatogonia are the only type of germ cells present in the seminiferous tubules during those developmental stages (Bellve http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Molecular Reproduction & Development Wiley

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Publisher
Wiley Subscription Services, Inc., A Wiley Company
Copyright
© 2018 Wiley Periodicals, Inc.
ISSN
1040-452X
eISSN
1098-2795
D.O.I.
10.1002/mrd.22961
Publisher site
See Article on Publisher Site

Abstract

AbbreviationsCRISPR/Cas9Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR‐associated‐9DMDDuchenne Muscular DystrophyDMEMDulbecco's Modified Eagle MediumDPBSDulbecco's PhoSphate Buffered SalineEDTAethylenediaminetetra acetic acidESCembryonic stem cellFACSfluorescence activated cell sortingGFPgreen fluorescent proteinHRhomologous recombinationiPSCinduced pluripotent stem cellMACSmagnetic activated cell sortingNHEJnon‐homologous end joiningSCNTsomatic cell nuclear transferSSCspermatogonial stem cellTALENTranscription Activator‐like Effector NucleaseUCH‐L1ubiquitin carboxyterminal hydrolase L‐1ZFNzinc finger nucleaseINTRODUCTIONSpermatogonia represent a diploid germ cell population in the seminiferous tubules which undergo mitotic division during spermatogenesis to give rise to primary spermatocytes. Spermatogonial stem cells (SSCs) are a subset of undifferentiated type A spermatogonia which have the capacity to self‐renew to maintain the SSC pool as well as to differentiate to ultimately form sperm. SSCs represent a rare cell population, constituting ∼0.03% of all germ cells in the adult mouse testis (Tegelenbosch & de Rooij, ). The lack of definite phenotypical, morphological, or biochemical markers that can unequivocally identify SSCs makes it impossible to isolate SSCs for in vitro studies. However, SSCs can be relatively enriched from a testicular cell suspension by various in vivo and/or in vitro approaches.For in vivo enrichment, neonatal, and prepubertal testes serve as the preferred source for harvesting germ cells as gonocytes/spermatogonia are the only type of germ cells present in the seminiferous tubules during those developmental stages (Bellve

Journal

Molecular Reproduction & DevelopmentWiley

Published: Jan 1, 2018

Keywords: ; ; ; ; ;

References

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