siRNA as a tool for investigating organogenesis: The pitfalls and the promises.
Removing the function of a specific gene from a developing organ, by making a 'knockout' mouse, is a powerful method for analyzing the molecular pathways that control organogenesis. The technique is expensive, though, in terms of time and money, and complex strategies for producing conditional knockouts are needed for genes that are essential for early development of the embryo, for which an unconditional knockout would be lethal before the organ of interest begins to form. Small interfering RNAs (siRNAs) offer a method of knocking down the expression of specific genes with no need for genomic manipulation. Almost as soon as they had been discovered, siRNAs began to be used to explore the molecular biology of mammalian cells in conventional, two-dimensional culture. They have now also been applied successfully, by several groups, to knock down specific genes in various organ rudiments developing in organ culture. This article reviews the basic technique of siRNA-mediated gene knockdown and how it is being applied to organ culture. It also reviews some of the current problems and challenges in the field, and the ways in which these problems are likely to be overcome.
Lee
Wen-Chin
WC
Centre for Integrative Physiology; University of Edinburgh; Edinburgh UK.
Berry
Rachel
R
Hohenstein
Peter
P
Davies
Jamie
J