Lentiviral Vector Delivery of siRNA and shRNA Encoding Genes into Cultured and Primary Hematopoietic Cells
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Lentiviral vectors are able to transduce non-dividing cells and maintain sustained long-term expression of the transgenes. Many cell types including brain, liver, muscle, and hematopoietic stem cells have been successfully transduced with lentiviral vectors carrying a variety of genes. These properties make lentiviral vectors attractive vehicles for delivering small interfering RNA (siRNA) genes into mammalian cells. RNA polymerase III (Pol III) promoters are most commonly used for expressing siRNAs from lentiviral vectors. Pol III promoters are relatively small, have high activity, and use simple termination signals of short stretches of U. It is possible to include several Pol III expression cassettes in a single lentiviral vector backbone to express different siRNAs or to combine siRNAs with other transgenes. This chapter describes the delivery of Pol III promoted siRNAs by HIV-based lentiviral vectors and covers vector design, production, and verification of siRNA expression and function. This chapter should be useful for establishing a lentiviral vector-based delivery of siRNAs in experiments that require long-term gene knockdown or developing siRNA-based approaches for gene therapy applications.
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- Lentiviral Vector Delivery of siRNA and shRNA Encoding Genes into Cultured and Primary Hematopoietic Cells
- Book Title
- Gene Therapy Protocols
- Book Subtitle
- Production and In Vivo Applications of Gene Transfer Vectors
- pp 287-300
- Print ISBN
- Online ISBN
- Series Title
- Methods in Molecular Biology™
- Series Volume
- Series ISSN
- Humana Press
- Copyright Holder
- Humana Press
- Additional Links
- Lentiviral vector
- RNA interference
- hematopoietic cells
- Industry Sectors
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