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Updated: May 13, 2025
Loss- and Gain-of-function Approach to Investigate Early Cell Fate Determinants in Preimplantation Mouse Embryos
Published on: June 6, 2016
Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection
Grace S Lee1, Natalie A Trigg2, Colin C Conine2
1Pharmacology 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, USA.
Abstract:
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.
