Hello.
I am planning an RNAi experiment.
I want to silence my gene of interest using RNAi.
My gene of interest is crucial for plant development.
Also, my gene of interest is known to function even when only c-terminal part is present in the cytoplasm. So what I'm thinking is, I could construct a vector that has partial DNA sequence for small interfering RNA and DNA fragments that would code C-terminal of my gene of interest.
I assume one transformant transformed with the vector containing only U6:RNAi sequences would show maldevelopment and the other transformed with the vector containing both U6:RNAi sequences and 35S:DNA fragment of gene of interest would show normal development.
I'm not sure my explanation was enough.
What I'm trying to do is silencing the target gene and doing transformation at the same time by using one vector (which contains siRNA sequences and half of DNA fragments of the target gene).
Hi Hyunbae Lee, It is adorable work; there is numerous publication on that.
In the attached article, they transform short hairpin RNA using the plasmid to silence the CXCR4 gene in the SW626 cancer cell line, it is a straight forward experiment.
Article Effect of short hairpin RNA-induced CXCR4 silence on ovarian...
It is theoretically feasible. But you need to pay attention to how to determine that RNAi has played a role. The rescue phenotype is indeed caused by the overexpressed c-terminal part.
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