rna interference: kill the messenger!. conclusion: dsrna triggers potent and specific gene silencing...
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RNA interference:
Kill the messenger!
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conclusion: dsRNA triggers potent and specific gene silencing
inject worms with dsRNAcorresponding to a geneinvolved in wiggling (unc-22)
Discovery
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Biochemistry to Examine Structure
short interfering RNA: siRNA
21 nt long, dsRNA:
P
P
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Dicer generates siRNAs
RNase III endonuclease
Mix cell extractwith dsRNA
21-mer
Berstein et al. Nature (2001)
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RNA interference (RNAi)dsRNA
siRNA
Dicer
RISC
AAAAcapmRNA
AAAAcapmRNA
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Timeline
1998
1999
2000
2001
2002
1990 cosuppression of purple color in plants
dsRNA injection in wormsshort RNAs identified in plants
RNAi shown in vitro
RISC activity partially purified
siRNAs identifiedDicer identified
RNAi used against HIVgenome-wide RNAi screens begin
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Big Potential for Small RNAs
Adapted from Ryther et.al. Gene Therapy (2005)
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S2 Drosophila cells spread on coated glass surface via lamellipodia
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Example of RNAi experiment