Gene transfection of lung cells in vitro and in vivo.

Abstract

This review focuses on the use of gene transfection as a tool with which to examine pulmonary function at the molecular, cellular, and whole organ level. The term transfection is defined as the introduction of one or more copies of a gene or cDNA into eukaryotic cells in order to permit either transient or permanent transcription and, usually, translation of a gene product, whose presence can be detected directly or inferred from functional assays. Although the term has also been applied to the introduction of DNA into embryonal cells to produce trans genic animals, this topic is not discussed in our review. Rather we describe various transfection techniques as well as experiments in which transfection-based methods have yielded new insights into pulmonary biology. We also discuss recent reports of successful in vivo lung cell transfection. In general, there are three reasons to consider gene transfection as an experimental approach. First, DNA can be transfected into a eukaryotic cell in order to produce a normal or mutated protein; the protein can then be characterized in terms of its structure and function in a physiologically relevant context. Two experiments utilizing the cystic fibrosis (CF) transmembrane conductance regulator (CFfR) gene illustrate this approach. In the first experiment, the CFfR cDNA was transfected into cells that lacked cAMP­ regulated chloride conductance (1 , 2); this ion transport capability is known to be absent or reduced in cells obtained from patients with CF. Both