Considering transitive RNAi and pre-mRNA targeting in C. elegans, does anyone know if there is a method of “isoform specific RNAi” in C. elegans?
Dear Hillel Schwartz
Thanks for your reply.
I had read the previous thread, but still it is not clear for me if there is a protocol for Isoform-Specific RNAi in C elegans. Therefore, I revived the subject again.
Here, I have a review of the previous thread:
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HillelSchwartz wrote: I would imagine this might be problematic in C.elegans…See the 2001 Sijen et al…
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The paper says: …the subsequent activity of DICER or another dsRNA-specific nuclease could function both (1) to destroy the mRNA and (2) to amplify the population of siRNA triggers.
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And: … Given the ability of the RdRP enzyme to initiate RNA synthesis at the end of a short RNA segment (Schiebel et al., 1993a, 1993b), it is certainly possible that the RdRP would carry out an additional reaction of copying sense segments of the input siRNA.
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cha osprod wrote:… knockdown of isoforms is possible. Murray et al…
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The paper says:
The distance between exon NÅå1 and exon 7 is ~ 36 kb. Exon NÅå3 was originally designated as exon 1 (Gross et al., 1990…
siRNA can be used to specifically target splice variants. We have shown for the first time in C. elegans that siRNAs corresponding to alternative first exons can specifically trigger the degradation of mRNA isoforms containing the targeted exon. Importantly, mRNA isoforms transcribed from the same gene, but which lack the targeted exon, are not degraded. We suggest that this strategy could be widely used in C. elegans to investigate the function of genes with alternative first exons.
I am not convinced yet. Does anyone have any suggestion?