使用显微注射评估精子RNA对小鼠着床前胚胎影响的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-06-20 Epub Date: 2025-05-09 DOI:10.1016/j.xpro.2025.103772

Grace S Lee, Natalie A Trigg, Colin C Conine

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引用次数: 0

摘要

受精卵显微注射表明，精子rna将非遗传表型传递给后代并影响胚胎发育。在这里，我们提出了一种微操作小鼠受精卵的方案，以引入生理相关水平的精子RNA。我们描述了在体外生产功能性mrna的步骤；纯化小鼠精子rna；准备，微注射，培养受精卵。该方案有助于精子RNA和受精后基因调控之间的因果分析。关于该协议的使用和执行的完整细节，请参考Trigg et al.1。

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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.¹.

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