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Questions on plant RNAi
Dear Colleagues,
I have several questions about practical plant RNAi to which I have been unable to find the answers in the literature. I suspect that these questions are mostly naive, but I since none of my colleagues here on my own campus work with plant RNAi, I thought I may be able to find the answers out here. First, are there any good practical guides or reviews out there on plant RNAi? Most of the literature that I have been able to dig up is on animal RNAi which is quite different in terms of RNAi probe design and activity. So far, the most practical advice has come from the Methods in Enzymology volume on RNAi. Second, again, there is a great deal of literature on designing the short RNAi probes for animals. There are even computer programs to help design those short sequences. However, I have been unable to come across any advice on choosing the long (300-500 bp) sequences needed for plant RNAi. Third, I am working with a family of plant genes, and I want to be able to knock down specific members of that family. What is the breakpoint percentage of homology at which you need no longer worry about cross-over inhibition against homologues? If the probe is 50% homologous to another family member which is not the target, will you get knockdown of the homologue? 40%? 30%? Is this even common knowledge, or does it come down to finding out these answers by practical experiments with your own species and gene family? Thanks for the help, Jim Campanella James J. Campanella, Associate Professor, Department of Biology and Molecular Biology Montclair State University 1 Normal Avenue Montclair, NJ 07043 Alternate email address: Ph: 973-655-4097 Fax: 973-655-7047 |
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