Cloning a transgene for transgenic RNAi in mouse oocytes
Language English Country United States Media print
Document type Journal Article
PubMed
20147029
DOI
10.1101/pdb.prot5134
PII: 2009/1/pdb.prot5134
Knihovny.cz E-resources
- MeSH
- Bacteria metabolism MeSH
- Cloning, Molecular * MeSH
- Molecular Sequence Data MeSH
- Mice, Transgenic MeSH
- Mice MeSH
- Oocytes cytology MeSH
- Plasmids metabolism MeSH
- Repetitive Sequences, Nucleic Acid MeSH
- RNA Interference * MeSH
- Base Sequence MeSH
- Transgenes * MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
RNA interference (RNAi) is a suitable method for sequence-specific post-transcriptional gene silencing for many model systems. The protocol presented here describes the cloning of a plasmid construct for use in transgenic RNAi experiments in mouse oocytes. The protocol is intended for production of a transgene by cloning an inverted repeat (IR) into the Zp3 transgenic cassette. The procedure begins with the selection of sequences and formulation of the cloning strategy. Subsequently, the IR is cloned, inserted into the transgenic cassette, and characterized by sequencing. Finally, the transgene is released from the cassette, purified, and provided to the transgenic facility.
References provided by Crossref.org
Shortcomings of short hairpin RNA-based transgenic RNA interference in mouse oocytes
