Retroviral delivery of ECM genes.
The use of recombinant DNA has become a powerful tool in the analysis of functional and structural properties of the extracellular matrix proteins. During last decade, various procedures of plasmid DNA delivery using liposome-based or electroporation-based transfection have been developed. However, in many instances, these procedures were shown to be not effective in DNA transfer or toxic for the mammalian cells. On contrary, retrovirus-mediated infection represents a superior mode of gene delivery with a success rate and viability of the cells approaching 100% in in vitro conditions. The use of the retroviral system also allows permanent insertion of the gene of interest into the chromosome of the infected cell, resulting in efficient gene transfer in which most recipient cells will incorporate and express the transduced gene. In this chapter, we will describe several retrovirus-based systems and provide step-by-step protocols applicable for the production of the recombinant virus and efficient delivery of the ECM genes.
Alexeev
Vitali
V
Department of Dermatology and Cutaneous Biology, Jefferson Medical College, Philadelphia, PA19107, USA. vitali.alexeev@jefferson.edu
Igoucheva
Olga
O
D004274:DNA, Recombinant / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D005822:Genetic Vectors; D012190:Retroviridae / Q000235:genetics*
D004274:DNA, Recombinant; D016326:Extracellular Matrix Proteins