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Genome editing in mouse spermatogonial stem/progenitor cells using engineered nucleases.


ABSTRACT: Editing the genome to create specific sequence modifications is a powerful way to study gene function and promises future applicability to gene therapy. Creation of precise modifications requires homologous recombination, a very rare event in most cell types that can be stimulated by introducing a double strand break near the target sequence. One method to create a double strand break in a particular sequence is with a custom designed nuclease. We used engineered nucleases to stimulate homologous recombination to correct a mutant gene in mouse "GS" (germline stem) cells, testicular derived cell cultures containing spermatogonial stem cells and progenitor cells. We demonstrated that gene-corrected cells maintained several properties of spermatogonial stem/progenitor cells including the ability to colonize following testicular transplantation. This proof of concept for genome editing in GS cells impacts both cell therapy and basic research given the potential for GS cells to be propagated in vitro, contribute to the germline in vivo following testicular transplantation or become reprogrammed to pluripotency in vitro.

SUBMITTER: Fanslow DA 

PROVIDER: S-EPMC4237364 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Genome editing in mouse spermatogonial stem/progenitor cells using engineered nucleases.

Fanslow Danielle A DA   Wirt Stacey E SE   Barker Jenny C JC   Connelly Jon P JP   Porteus Matthew H MH   Dann Christina Tenenhaus CT  

PloS one 20141119 11


Editing the genome to create specific sequence modifications is a powerful way to study gene function and promises future applicability to gene therapy. Creation of precise modifications requires homologous recombination, a very rare event in most cell types that can be stimulated by introducing a double strand break near the target sequence. One method to create a double strand break in a particular sequence is with a custom designed nuclease. We used engineered nucleases to stimulate homologou  ...[more]

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