Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection
Summary: Zygotic microinjections show that sperm RNAs transmit non-genetically inherited phenotypes to offspring and influence embryonic development. Here, we present a protocol for the micromanipulation of mouse zygotes to introduce physiologically relevant levels of sperm RNA. We describe steps fo...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725001789 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849729482347249664 |
|---|---|
| author | Grace S. Lee Natalie A. Trigg Colin C. Conine |
| author_facet | Grace S. Lee Natalie A. Trigg Colin C. Conine |
| author_sort | Grace S. Lee |
| collection | DOAJ |
| description | Summary: Zygotic microinjections show that sperm RNAs transmit non-genetically inherited phenotypes to offspring and influence embryonic development. Here, we present a protocol for the micromanipulation of mouse zygotes to introduce physiologically relevant levels of sperm RNA. We describe steps for producing functional mRNAs in vitro; purifying mouse sperm RNAs; and preparing, microinjecting, and culturing zygotes. This protocol facilitates causal analysis between sperm RNA and gene regulation postfertilization.For complete details on the use and execution of this protocol, please refer to Trigg et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-ba07f8a12a784c779a762f529e5934d5 |
| institution | DOAJ |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-ba07f8a12a784c779a762f529e5934d52025-08-20T03:09:12ZengElsevierSTAR Protocols2666-16672025-06-016210377210.1016/j.xpro.2025.103772Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjectionGrace S. Lee0Natalie A. Trigg1Colin C. Conine2Pharmacology Graduate Group, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Departments of Genetics and Pediatrics - Penn Epigenetics Institute, Institute of Regenerative Medicine, and Center for Women’s Health and Reproductive Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USADivision of Neonatology, Children’s Hospital of Philadelphia, Philadelphia, PA, USA; Departments of Genetics and Pediatrics - Penn Epigenetics Institute, Institute of Regenerative Medicine, and Center for Women’s Health and Reproductive Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USADivision of Neonatology, Children’s Hospital of Philadelphia, Philadelphia, PA, USA; Departments of Genetics and Pediatrics - Penn Epigenetics Institute, Institute of Regenerative Medicine, and Center for Women’s Health and Reproductive Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA; Corresponding authorSummary: Zygotic microinjections show that sperm RNAs transmit non-genetically inherited phenotypes to offspring and influence embryonic development. Here, we present a protocol for the micromanipulation of mouse zygotes to introduce physiologically relevant levels of sperm RNA. We describe steps for producing functional mRNAs in vitro; purifying mouse sperm RNAs; and preparing, microinjecting, and culturing zygotes. This protocol facilitates causal analysis between sperm RNA and gene regulation postfertilization.For complete details on the use and execution of this protocol, please refer to Trigg et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725001789Developmental biologyGene ExpressionBiotechnology and bioengineering |
| spellingShingle | Grace S. Lee Natalie A. Trigg Colin C. Conine Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection STAR Protocols Developmental biology Gene Expression Biotechnology and bioengineering |
| title | Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection |
| title_full | Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection |
| title_fullStr | Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection |
| title_full_unstemmed | Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection |
| title_short | Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection |
| title_sort | protocol to assess the impact of sperm rna on mouse preimplantation embryos using microinjection |
| topic | Developmental biology Gene Expression Biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725001789 |
| work_keys_str_mv | AT graceslee protocoltoassesstheimpactofspermrnaonmousepreimplantationembryosusingmicroinjection AT natalieatrigg protocoltoassesstheimpactofspermrnaonmousepreimplantationembryosusingmicroinjection AT colincconine protocoltoassesstheimpactofspermrnaonmousepreimplantationembryosusingmicroinjection |