This month’s issue of Cold Spring Harbor Protocols highlights two methods to understand developmental processes in plants and flies. Both methods involve work with RNA and are freely accessible on the Web site for Cold Spring Harbor Protocols.
The first protocol describes the use of RNAi to investigate gene function in fruit flies. Short RNA molecules are injected into fly embryos to disrupt the function of a specific gene. The embryos are then allowed to grow until they have reached a desired stage of development, when they are evaluated for changes that may have occurred due to the disruption of the gene.
The protocol, freely accessible at www.cshprotocols.org/cgi/content/full/2008/3/pdb.prot4918, is a recently updated version of one that originally appeared in the book Drosophila Protocols. It was developed by Dr. Bruce Paterson’s group at the National Institutes of Health. His lab has used the protocol to investigate the regulation of gene expression during development, especially as it relates to muscle formation in the embryo.
The second protocol, available at www.cshprotocols.org/cgi/content/full/2008/3/pdb.prot4944, describes how to detect where and when a gene is expressed in young plant tissues such as embryos, seedlings, floral tissues, and other developing organs. The tissues are stained with a non-radioactive probe that specifically targets RNA molecules from a gene of interest. The staining pattern is then detected by microscopy.
Scientists can analyze this staining pattern in a variety of tissue samples at different stages in development to gain a better understanding of the function of that gene. The method can be used to study virtually any plant species and will be useful to many biologists.