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Status |
Public on Aug 01, 2022 |
Title |
Generation of functional rat spermatozoa in sterile mice utilizing blastocyst complementation with pluripotent stem cells [scRNAseq_rat] |
Organism |
Rattus norvegicus |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
Pluripotent stem cells (PSCs) provide a powerful tool to produce transgenic animals for biomedical research. However, impaired PSC contribution to chimerism and most notably to the germline oftentimes impedes production of genetically modified animals, rendering techniques that expediate PSC contribution to the germline highly desirable. Blastocyst complementation denotes a technique which purposes to generate organs, tissues or cells in animal chimeras following microinjection of PSCs into genetically compromised blastocyst-stage embryos. Here, we report on successful blastocyst complementation of the male germline in adult chimeras via microinjection of mouse or rat PSCs into mouse blastocysts mutated for Tsc22d3, an essential gene for spermatozoa production. Microinjection of mouse embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) into Tsc22d3-KnockOut (KO) blastocyst-stage embryos gave rise to intraspecies chimeras embodying functional spermatozoa, which were solely derived from the microinjected PSCs. Furthermore, microinjection of rat ESCs into Tsc22d3-KO mouse embryos gave rise to viable mouse-rat chimeras that exhibited extensive contribution of rat cells to various tissues and organs. Notably, multiple mouse-rat chimeras showed contribution of rat ESCs to the male germline, solely harboring rat spermatids and spermatozoa that were rat ESC-derived and could fertilize rat oocytes. Collectively, this study reports on a method for exclusive germ cell production of one species in another via blastocyst complementation with PSCs. Implications of this study may extend to development of transgenic rat gametes in sterile mice and could further assist efforts to generate germ cells from endangered animal species.
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Overall design |
Samples were isolated from an adult mouse-rat chimera testis (host: goGermline, Ozgene. Donor: DAC8 rat ESCs) via the protocol published by Valli et al, Fertility and Sterility, 2014
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Contributor(s) |
Zvick J, Ghosh A, Bar-Nur O |
Citation(s) |
35931077 |
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Submission date |
May 19, 2022 |
Last update date |
Oct 04, 2022 |
Contact name |
Ori Bar-Nur |
E-mail(s) |
ori.bar-nur@hest.ethz.ch
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Organization name |
ETH Zurich
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Department |
Health Sciences and Technology
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Lab |
Regenerative and Movement Biology
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Street address |
Schorenstrasse 16
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City |
Schwerzenbach |
State/province |
Zurich |
ZIP/Postal code |
8603 |
Country |
Switzerland |
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Platforms (1) |
GPL25947 |
Illumina NovaSeq 6000 (Rattus norvegicus) |
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Samples (1) |
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This SubSeries is part of SuperSeries: |
GSE167435 |
Generation of functional rat spermatozoa in sterile mice utilizing blastocyst complementation with pluripotent stem cells |
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Relations |
BioProject |
PRJNA839614 |