RNAi Mouse Embryos and Oocytes Protocols

Protocol describes a method to collect early embryos from 6-week-old mice.  Subsequently, the isolated embryos can be injected with double-stranded RNA to induce knockdown of a gene of interest. - [Read Collection of Early Mouse Embryos for RNAi Protocol]

Protocol describes a method to collect oocytes from 6-week-old mice.  Subsequently, the isolated oocytes can be injected with double-stranded RNA to induce knockdown of a gene of interest. - [Read Collection of Mouse Oocytes for RNAi Protocol]

Protocol describes how to introduce a double-stranded RNA (dsRNA) of choice into mouse oocytes or fertilized one-cell embryos by microinjection.  For collection of mouse oocytes and early embryos, see Collection of Mouse Oocytes for RNAi and Collection of Early Mouse Embryos for RNAi. - [Read Microinjection of dsRNA into Mouse Oocytes and Early Embryos Protocol]

Protocol describes in vitro transcription with the SP6 polymerase.  For other polymerases (T3, T7), simply use the appropriate buffer and desired RNA polymerase instead of the SP6 polymerase. - [Read Preparation of dsRNA Molecules for RNAi in Mouse Oocytes and Early Embryos Protocol]

Protocol describes the use of RT-PCR to confirm knockdown of a gene of interest in RNAi-treated mouse oocytes and early embryos.  All of the solutions used during RNA isolation and reverse transcription must be Rnase-free - [Read RNA Isolation and RT-PCR from dsRNA-Treated Mouse Oocytes and Early Embryos Protocol]