将双链RNA显微注射到小鼠单细胞胚胎中。

Microinjection of dsRNA into mouse one-cell embryos.

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.prot5133. doi: 10.1101/pdb.prot5133.

DOI:10.1101/pdb.prot5133

PMID:20147028

Abstract

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 mouse one-cell embryos by microinjection. Microinjected embryos can be cultured for up to 4 d, i.e., to the blastocyst stage. The efficiency of knockdown by RNAi can be assayed by quantitative reverse transcriptase (RT)-PCR (qPCR).

摘要

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选择的双链RNA（dsRNA）导入小鼠单细胞胚胎的过程。显微注射后的胚胎可以培养长达4天，即到囊胚阶段。RNAi敲低的效率可以通过定量逆转录酶（RT）-PCR（qPCR）进行检测。