使用显微注射评估精子RNA对小鼠植入前胚胎影响的实验方案。

Protocol to assess the impact of sperm RNA on mouse preimplantation embryos using microinjection.

作者信息

Lee Grace S, Trigg Natalie A, Conine Colin C

机构信息

Pharmacology 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.

Division 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.

出版信息

STAR Protoc. 2025 Jun 20;6(2):103772. doi: 10.1016/j.xpro.2025.103772. Epub 2025 May 9.

DOI:10.1016/j.xpro.2025.103772

PMID:40347477

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12136814/

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..

摘要

合子显微注射表明，精子RNA可将非遗传继承的表型传递给后代，并影响胚胎发育。在此，我们提供了一个用于小鼠合子显微操作的方案，以引入生理相关水平的精子RNA。我们描述了体外产生功能性mRNA、纯化小鼠精子RNA以及制备、显微注射和培养合子的步骤。该方案有助于对受精后精子RNA与基因调控之间进行因果分析。有关该方案使用和执行的完整详细信息，请参考特里格等人的研究。