Lentiviral vector delivery of recombinant small interfering RNA expression cassettes

Ming Jie Li, John J. Rossi

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

Lentiviral vectors are able to transduce nondividing cells and maintain sustained long-term expression of transgenes. Many cells 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 Us. 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 human immunodeficiency virus (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.

Original languageEnglish
Pages (from-to)218-226
Number of pages9
JournalMethods in enzymology
Volume392
DOIs
StatePublished - Jan 24 2005
Externally publishedYes

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