将双链RNA显微注射到完全成熟的小鼠卵母细胞中。
Microinjection of dsRNA into fully-grown mouse oocytes.
作者信息
Stein Paula
机构信息
Department of Biology, University of Pennsylvania, Philadelphia, PA 19104, USA.
出版信息
Cold Spring Harb Protoc. 2009 Jan;2009(1):pdb.prot5132. doi: 10.1101/pdb.prot5132.
RNA interference (RNAi) is a suitable method for sequence-specific post-transcriptional gene silencing in a number of model systems. The following protocol describes delivery of a double-stranded RNA (dsRNA) of choice into fully-grown, germinal vesicle-intact (GV) mouse oocytes by microinjection. Microinjected oocytes can be cultured for up to 2 d or they can be matured to metaphase II. The metaphase II eggs can be fertilized in vitro and cultured up to the blastocyst stage. The efficiency of knockdown by RNAi can be assayed by quantitative reverse transcriptase (RT)-PCR (qPCR).
RNA干扰（RNAi）是在许多模型系统中进行序列特异性转录后基因沉默的合适方法。以下方案描述了通过显微注射将所选双链RNA（dsRNA）导入完全成熟、生发泡完整（GV）的小鼠卵母细胞。显微注射的卵母细胞可以培养长达2天，或者可以使其成熟至中期II。中期II期的卵可以在体外受精并培养至囊胚阶段。RNAi的敲低效率可以通过定量逆转录酶（RT）-PCR（qPCR）进行检测。
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