Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells
Organisms with targeted genomic modifications are efficiently produced by gene editing in embryos using CRISPR/Cas9 RNA-guided DNA endonuclease. Here, to facilitate germline editing in rats, we used CRISPR/Cas9 to catalyze targeted genomic mutations in rat spermatogonial stem cell cultures. CRISPR/C...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2015-03-01
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| Series: | Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124715001989 |
| _version_ | 1819210234126663680 |
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| author | Karen M. Chapman Gerardo A. Medrano Priscilla Jaichander Jaideep Chaudhary Alexandra E. Waits Marcelo A. Nobrega James M. Hotaling Carole Ober F. Kent Hamra |
| author_facet | Karen M. Chapman Gerardo A. Medrano Priscilla Jaichander Jaideep Chaudhary Alexandra E. Waits Marcelo A. Nobrega James M. Hotaling Carole Ober F. Kent Hamra |
| author_sort | Karen M. Chapman |
| collection | DOAJ |
| description | Organisms with targeted genomic modifications are efficiently produced by gene editing in embryos using CRISPR/Cas9 RNA-guided DNA endonuclease. Here, to facilitate germline editing in rats, we used CRISPR/Cas9 to catalyze targeted genomic mutations in rat spermatogonial stem cell cultures. CRISPR/Cas9-modified spermatogonia regenerated spermatogenesis and displayed long-term sperm-forming potential following transplantation into rat testes. Targeted germline mutations in Epsti1 and Erbb3 were vertically transmitted from recipients to exclusively generate “pure,” non-mosaic mutant progeny. Epsti1 mutant rats were produced with or without genetic selection of donor spermatogonia. Monoclonal enrichment of Erbb3 null germlines unmasked recessive spermatogenesis defects in culture that were buffered in recipients, yielding mutant progeny isogenic at targeted alleles. Thus, spermatogonial gene editing with CRISPR/Cas9 provided a platform for generating targeted germline mutations in rats and for studying spermatogenesis. |
| first_indexed | 2024-12-23T06:07:56Z |
| format | Article |
| id | doaj.art-3636e9614fd349e99ff5d4fa5d2528d2 |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-12-23T06:07:56Z |
| publishDate | 2015-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-3636e9614fd349e99ff5d4fa5d2528d22022-12-21T17:57:32ZengElsevierCell Reports2211-12472015-03-0110111828183510.1016/j.celrep.2015.02.040Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem CellsKaren M. Chapman0Gerardo A. Medrano1Priscilla Jaichander2Jaideep Chaudhary3Alexandra E. Waits4Marcelo A. Nobrega5James M. Hotaling6Carole Ober7F. Kent Hamra8Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USADepartment of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USADepartment of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USADepartment of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USADepartment of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USADepartment of Human Genetics, University of Chicago, Chicago, IL 60637, USADepartment of Surgery (Urology), University of Utah School of Medicine, Salt Lake City, UT 84134, USADepartment of Human Genetics, University of Chicago, Chicago, IL 60637, USADepartment of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USAOrganisms with targeted genomic modifications are efficiently produced by gene editing in embryos using CRISPR/Cas9 RNA-guided DNA endonuclease. Here, to facilitate germline editing in rats, we used CRISPR/Cas9 to catalyze targeted genomic mutations in rat spermatogonial stem cell cultures. CRISPR/Cas9-modified spermatogonia regenerated spermatogenesis and displayed long-term sperm-forming potential following transplantation into rat testes. Targeted germline mutations in Epsti1 and Erbb3 were vertically transmitted from recipients to exclusively generate “pure,” non-mosaic mutant progeny. Epsti1 mutant rats were produced with or without genetic selection of donor spermatogonia. Monoclonal enrichment of Erbb3 null germlines unmasked recessive spermatogenesis defects in culture that were buffered in recipients, yielding mutant progeny isogenic at targeted alleles. Thus, spermatogonial gene editing with CRISPR/Cas9 provided a platform for generating targeted germline mutations in rats and for studying spermatogenesis.http://www.sciencedirect.com/science/article/pii/S2211124715001989 |
| spellingShingle | Karen M. Chapman Gerardo A. Medrano Priscilla Jaichander Jaideep Chaudhary Alexandra E. Waits Marcelo A. Nobrega James M. Hotaling Carole Ober F. Kent Hamra Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells Cell Reports |
| title | Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells |
| title_full | Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells |
| title_fullStr | Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells |
| title_full_unstemmed | Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells |
| title_short | Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells |
| title_sort | targeted germline modifications in rats using crispr cas9 and spermatogonial stem cells |
| url | http://www.sciencedirect.com/science/article/pii/S2211124715001989 |
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