Biology Forum › Molecular Biology › RNA interference
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- August 18, 2010 at 6:04 am #13664
tgkParticipantIn plants for getting a hairpin loop structure in pHannibal vector people normally use cDNA as the starting material from which to amplify the gene of interest. My question is whether one can start with the genomic DNA? What happens if introns are present in the region which will form ds RNA after transcription? The basic idea being that cDNA preparation can be avoided.
- August 19, 2010 at 5:58 am #100911JackBeanParticipant
Hi,
there are several modes of RNA interference, one of them is the binding to DNA thus blocking transcription. Unfortunately, I don’t remember, whether this is plant’s or animal’s case…
If it was possible, then using gDNA would be fine. If it worked classicaly through binding to mRNA and it’s digest, then your RNAi would have non-complementar part and probably would not work 😉 - August 19, 2010 at 9:35 am #100920tgkParticipant
Thank you. You are right. Since the interference is by binding of antisense RNA strand of 21-24 nt in RISC to mRNA, if introns are present they will not have complementary sequence in mRNA. In short, there is no short cut. It has to be cDNA.
- August 19, 2010 at 11:41 am #100922JackBeanParticipant
well, I didn’t say exactly that, but you’re rigth
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