Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection
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.<sup>1</sup>.
Authors
- Colin C. Conine (ORCID: https://orcid.org/0000-0001-7915-0463)
- Grace S. Lee (ORCID: https://orcid.org/0000-0003-0329-7906)
- Natalie A. Trigg (ORCID: https://orcid.org/0000-0002-8999-5731)
Institutions
- Children's Hospital of Philadelphia (US)
- University of Pennsylvania (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-05-09
- DOI
- https://doi.org/10.1016/j.xpro.2025.103772
- Citations
- 3
- Primary Topic
- Molecular Biology Techniques and Applications
- Type
- article
- Field-Weighted Citation Impact
- 2.42
Funders
- Pew Charitable Trusts
- Lalor Foundation
- National Institute of Child Health and Human Development
- Eunice Kennedy Shriver National Institute of Child Health and Human Development