Chapter 19 DNA Transfection of Cultured Muscle Cells

Abstract

Publisher Summary This chapter discusses the DNA transfection of cultured muscle cells. Myoblasts are fairly straightforward to isolate from embryonic and neonatal muscles, and myogenic cultures are generally amenable to transfection with exogenous DNA. Assays of gene expression from these transient DNA structures are most useful when rapid testing of numerous constructs is required, as multiple transient transfections can be harvested in parallel. The popularity of muscle cell culture systems for the analysis of cloned gene expression lies with the standard techniques involved and with the reproducibility of the assays. Following introduction of expression vector DNA into myoblast cell cultures, transient expression of the exogenous genes can usually be measured for up to 72 hr post-transfection. The virtues of transient assays include the convenience of the short time between transfection and harvest. This is ideal for the design of expression studies such as deletion or mutation analysis of a DNA regulatory sequence, requiring sequential testing of many constructs.