siRNA-mediated knockdown of a splice variant of the PK-A catalytic subunit gene causes adult-onset paralysis in C. elegans |
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Authors: | Murray Patricia Clegg Roger A Rees Huw H Fisher Michael J |
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Affiliation: | School of Biological Sciences, Biosciences Building, University of Liverpool, Liverpool, UK. p.a.murray@liv.ac.uk |
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Abstract: | In C. elegans, the PK-A catalytic subunit is encoded by kin-1, which has six 5' exons (N'1-N'6), any one of which may be alternatively spliced onto exon-2. Here we describe a novel siRNA-based strategy to knockdown the expression levels of the N'3 and N'4 splice variants. We show that this technique can effectively knockdown expression of the targeted isoforms without affecting expression of the other kin-1 splice variants. We suggest that this strategy could be widely used in C. elegans to investigate the function of genes with alternative first exons. Moreover, we report a novel role for the N'3 kin-1 variant. Whereas knockdown of the N'4 variant results in no obvious phenotype, loss of the N'3 variant leads to paralysis and an egg-laying defect in the adult, suggesting a deficit in the function of the neuromuscular junction. The function of the N'3 variant is discussed in relation to the known function of PK-A in regulation of the release of neurotransmitters from many presynaptic termini. |
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