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BISC219/F12: RNAi Lab 7

No change in size, 12:17, 12 November 2012
Lab 7: Series 3-Reverse Genetics: Investigating Gene Function & Regulation Using RNAi
We are going to use RNAi as our tool to investigate gene function via reverse genetics. ''C. elegans'' is the first animal in which the process of RNAi was discovered. A similar system was identified in plants years earlier but, curiously, that groundbreaking discovery was largely ignored by the scientific community until it was noticed in animal models. We now know that RNA regulation in cells is a fundamental method of regulating gene expression in organisms ranging from microscopic ''C. elegans'' to humans. Many labs are now working non-stop to develop treatments for many "incurable" human diseases using RNAi.<br><Br>
In this investigation for Series 3 that you will begin today, each pair of students will be given ''E. coli'' bacteria that have been genetically manipulated to contain a plasmid that contains the ''C. elegans'' ''lsy-2'' gene. The bacteria that contain this plasmid express ''C. elegans lyslsy-2'' double stranded RNA. These bacteria will serve as food for our wild type ''C. elegans'' (worms with normal copies of ''lsy-2'') and induce the RNA interference pathway in those worms. The result should be a '"knock down" in the amount of mRNA specific to the ''lsy-2'' gene and, thus, a reduction in the amount of ''lsy-2'' gene product. We hope to see the effect of this gene silencing in phenotypic differences between wild type worms that were RNAi induced and those that were not.<br><br>
== Calibration of Micropipettes ==
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