Protocol

Lentivirus Transduction of Hematopoietic Cells

This protocol was adapted from "Lentivirus Transduction of Hematopoietic Cells," Chapter 7, in Gene Transfer: Delivery and Expression of DNA and RNA (eds. Friedmann and Rossi). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2007.

INTRODUCTION

Efficient transfer and sustained expression of transgenes are among the most important issues in gene delivery. The majority of hematopoietic cells are nondividing or slowly self-renewing. Thus, they are refractory to most nonviral or retroviral delivery methods. Lentiviral vectors are capable of transducing nondividing cells and maintaining long-term and sustained expression of the transgenes. They are becoming useful for many delivery protocols, such as long-term expression of short hairpin RNA (shRNA) and functional genetics. They may also have great potential in gene therapy. This protocol describes lentivirus-vector-based delivery of foreign genes to hematopoietic cells. The method is applicable to various cell types in experiments that require long-term transgene expression.