Examining the Rna Interference Mechanism in the Dpy-13 Gene in C. Elegans Through Feeding

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Examining the RNA Interference Mechanism in the dpy-13 Gene in C. Elegans Through Feeding
Mehdi Misto
Lab: Monday 1:00 – 4:50 PM
11 December 2012

Introduction: RNA interference, or RNAi, is a biological process in which RNA molecules reduce the gene expression of an organism. This is done typically by causing the destruction of specific mRNA molecules. RNAs are direct products of genes, these small RNAs can bind to other mRNA molecules to either increase or decrease their activity like in the example of preventing an mRNA from producing a protein. There are two types of RNA molecules that are central to RNAi, these molecules are, micro RNA (miRNA) and small interfering RNA (siRNA).
The RNAi mechanism is found in many different
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This same procedure is repeated to transfer worms from the dpy-13 starter plate to the OP50 seeded plate labeled “dpy-13”. These plates are then both incubated upside down at 20˚C for 48 hours. This incubation gives the worms time to grow in their new environment. In part 2 of the experiment we are inducing RNAi through the process of feeding. To do so, a black pen is used to label the bottom of an OP50 seeded plate with the date and “wild type”. Another OP50 seeded plate is labeled with the date and “dpy-13”. A black pen is then used to label the bottom of the plate seeded with dpy-13 RNAi feeding strain with the date and “wild type”. Five L4-stage worms from the plate of wild type worms are then picked and moved to the OP50 seeded plate labeled “wild type”. Any eggs or young larvae that may have been accidentally transferred are to be picked off of the plate and then flamed in a Bunsen burner. The same method is used to move five L4 wild type worms to the plate seeded with the dpy-13 RNAi feeding strain, and once again to move five L4 dpy-13 worms to the OP50 seeded plate labeled “dpy-13”. These plates are then incubated upside down at 20˚C. Part 3 of this experiment calls to isolate the DNA from C. Elegans. To do so, we start by labeling three PCR tubes with the group number. One of the tubes is labeled “W” for wild type, one is labeled “R” for

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