What are the pros and cons of using RNAi vs. creating a knockout?
I guess RNAi would be more convenient/easier in most cases...
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RNAi vs. knockouts
Started by
wraith
, May 13 2005 04:14 PM
4 replies to this topic
#3
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Posted 13 May 2005 - 11:40 PM
One criterion that should affect your choice is that with RNAi you often knock the gene product down to some 20% but do not eliminate it like with a ko. (In C. elegans it seems to be somewhat more effective than in other animals)
#4
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Posted 14 May 2005 - 05:14 AM
guess RNAi would be more convenient/easier in most cases...
Correct. But like John said, it does not give you 100% suppression of gene expression like a knockout does. Another benefit is that you use a promoter of your choice to regulate expression, eg you can localize the silencing effect by using a tissue specific promoter (or simply by direct injection to the region you wish to silence). What is the organism you are working/looking to work with?
This is a very versatile and powerful technology whose implications are only beginning to be understood. For instance, Dicer processed dsRNA not only acts post-transcriptionally to suppress gene expression by hybridising to the homologous mRNA region but it also acts on a DNA chromatin methylation level to silence expression pre-transcriptionally. Futhermore, miRNAs are now accepted as being another level of gene regulation where ssRNA self-hybridizes forming a hairpin structure which upon Dicer processing also supresses translation of mRNA. Such a diversity of RNAi endogenous gene supression mechanisms (originally RNAi was thought to act only as a defence against retroviruses) suggests that we have evolved a lot machinery to regulate expression and consequently presents a whole new range of intervention opportunities, from chromatin bound DNA to mRNA.
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