Rapid generation of knockdown transgenic mice by silencing lentiviral vectors.
Lentiviral vectors are potent gene delivery vehicles that enable stable expression of transgenes in both dividing and postmitotic cells, including preimplantation embryos. We have developed lentiviral vectors carrying silencing cassettes consisting of an RNA polymerase III promoter expressing short hairpin RNAs. Transgenic mice can be generated rapidly by transduction of early embryos with lentiviral silencing vectors, resulting in mice with downregulated target genes. We describe two alternative early embryo transduction protocols (removal of zona pellucida and subzonal microinjection). These methodologies offer the possibility of large-scale generation of knockdown transgenic mice for functional genomic studies and enable the production of transgenic mice in 7 weeks.
Singer
Oded
O
The Salk Institute for Biological Studies, Laboratory of Genetics, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.
Tiscornia
Gustavo
G
Ikawa
Masahito
M
Verma
Inder M
IM
D000818:Animals; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic / Q000196:embryology / Q000235:genetics*; D008846:Micromanipulation; D034622:RNA Interference; D014161:Transduction, Genetic / Q000379:methods*