Targeted Germline Modifications in Rats Using CRISPR/Cas9 and Spermatogonial Stem Cells

Cell Reports
Karen M ChapmanF Kent Hamra

Abstract

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.

References

Jun 16, 1989·Science·V G PurselR E Hammer
Jan 26, 1989·Nature·R E BraunR D Palmiter
Nov 1, 1986·Developmental Biology·T M WilkieR D Palmiter
Nov 22, 1994·Proceedings of the National Academy of Sciences of the United States of America·R L Brinster, M R Avarbock
Apr 25, 2001·Biology of Reproduction·M NaganoR L Brinster
Oct 19, 2001·Proceedings of the National Academy of Sciences of the United States of America·M NaganoR L Brinster
Oct 23, 2002·Proceedings of the National Academy of Sciences of the United States of America·F Kent HamraDavid L Garbers
Jun 3, 1961·Nature·A K TARKOWSKI
May 25, 2004·Nature Genetics·F William BuaasRobert E Braun
May 25, 2004·Nature Genetics·José A CostoyaPier Paolo Pandolfi
Oct 1, 2004·Biology of Reproduction·Mito Kanatsu-ShinoharaTakashi Shinohara
Jun 7, 2005·Nature·Yasuhide OhinataM Azim Surani
Nov 19, 2005·Proceedings of the National Academy of Sciences of the United States of America·F Kent HamraDavid L Garbers
Nov 14, 2006·The Journal of Biological Chemistry·F Kent HamraDavid L Garbers
May 18, 2010·Nature Methods·Zsuzsanna IzsvákF Kent Hamra
Oct 1, 2011·Nature Protocols·Zoltán IvicsF Kent Hamra
May 29, 2012·American Journal of Human Genetics·Gülüm KosovaCarole Ober
Jan 5, 2013·Science·Le CongFeng Zhang
Aug 10, 2013·Nature Biotechnology·Dali LiMingyao Liu
May 6, 2014·Annales d'endocrinologie·Denise R Archambeault, Martin M Matzuk
Jul 2, 2014·Developmental Biology·Shuo-Ting YenRichard R Behringer
Sep 4, 2014·Genes & Development·Melissa M HarrisonJill Wildonger

❮ Previous
Next ❯

Citations

Jan 10, 2016·Fertility and Sterility·Kathrin Gassei, Kyle E Orwig
Jan 13, 2016·Cellular and Molecular Life Sciences : CMLS·Yan ZhouYonglun Luo
Mar 5, 2016·Transgenic Research·Teppei GotoMasumi Hirabayashi
Apr 26, 2016·Molecular Therapy. Nucleic Acids·Bhola Shankar Pradhan, Subeer S Majumdar
Nov 19, 2015·Biological Procedures Online·A R GrassianS Ribich
Oct 4, 2015·Trends in Molecular Medicine·Lukas E Dow
May 18, 2016·Neurobiology of Disease·Maryna KoskelaMikko Airavaara
Mar 10, 2016·Asian-Australasian Journal of Animal Sciences·Wanyou FengDeshun Shi
Dec 3, 2016·Regenerative Medicine·Kuang-Yui Chen, Paul S Knoepfler
Jan 6, 2017·Biotechnology & Genetic Engineering Reviews·Amir Roointan, Mohammad Hossein Morowvat
Jan 18, 2017·Genesis : the Journal of Genetics and Development·Paula G SlaterLaura Anne Lowery
Mar 1, 2017·Nature Methods·Johannes Bohacek, Isabelle M Mansuy
Jun 4, 2017·Reproduction : the Official Journal of the Society for the Study of Fertility·Yi ZhengGeert Hamer
Nov 15, 2017·Reproduction : the Official Journal of the Society for the Study of Fertility·Go Nagamatsu, Katsuhiko Hayashi
Oct 23, 2015·Cell Death & Disease·J Chaudhary, F K Hamra
Mar 4, 2017·Circulation Research·Thomas Doetschman, Teodora Georgieva
Feb 7, 2019·In Vitro Cellular & Developmental Biology. Animal·Min Seong KimSeung Tae Lee
Feb 21, 2019·The Journal of Gene Medicine·Ali ZareiSeyed Mohammad Bagher Tabei
Jun 18, 2019·Journal of Animal Science and Biotechnology·Filipp SavvulidiLudek Stadnik
Mar 4, 2020·Briefings in Functional Genomics·Min XuJiansong Ji
Mar 13, 2020·Human Reproduction Update·Murat GulEllen Goossens
Jul 7, 2017·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·András TálasErvin Welker
Apr 28, 2020·Cell and Tissue Research·Fahar IbtishamAli Honaramooz
Feb 1, 2018·Biology of Reproduction·Ewelina Bolcun-Filas, Mary Ann Handel
Mar 25, 2017·Biology of Reproduction·Kathrin GasseiKyle E Orwig
Feb 3, 2018·Molecular Reproduction and Development·Lin TangIna Dobrinski
Nov 1, 2018·Reproductive Medicine and Biology·Seiji Takashima
Sep 19, 2018·Therapeutic Innovation & Regulatory Science·Gregory Katz, Peter J Pitts
Aug 26, 2020·Proceedings of the National Academy of Sciences of the United States of America·Yike YinZhonghan Li
Feb 2, 2019·Scientific Reports·Leila GhanbariSuhasa B Kodandaramaiah
Apr 5, 2018·Biology of Reproduction·Hiroshi Kubota, Ralph L Brinster
Feb 8, 2018·Biologie aujourd'hui·Pierre Jouannet
Feb 16, 2021·Frontiers in Genetics·Dennis WebsterIna Dobrinski
May 1, 2021·Frontiers in Genetics·Jason C Lin, Alison L Van Eenennaam
Jun 3, 2021·Genes·Ovidiu BîcăCarmen Iulia Ciongradi

❮ Previous
Next ❯

Related Concepts

Related Feeds

Adult Stem Cells

Adult stem cells reside in unique niches that provide vital cues for their survival, self-renewal, and differentiation. They hold great promise for use in tissue repair and regeneration as a novel therapeutic strategies. Here is the latest research.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.