Abstract
A fitness assay for comparing RNAi effects across multiple C. elegans genotypes
Mark Elvin¹, Basten L Snoek², Martin Frejno¹, Ulrike Klemstein¹, Jan E Kammenga², Gino B Poulin¹
1University of Manchester, Faculty of Life Sciences, Manchester, M13 9PT, United Kingdom,
2Wageningen University, Nematology, Wageningen, 7608PB, Netherlands
RNAi technology by feeding C. elegans with E. coli containing dsRNA has significantly
contributed to increase our understanding of many different fields of biology, including genetics,
molecular biology, developmental biology and functional genomics. Most of this research has
been carried out in a single genotype or genetic background. However, RNAi effects in one
genotype do not reveal the allelic effects that segregate in natural populations and contribute to
phenotypic variation.
Here we present a method that allows for rapidly comparing RNAi effects among diverse
genotypes at an improved high throughput rate. It is based on assessing the fitness of a
population of worms by measuring the rate at which E. coli is consumed, the feeding curve.
Critically, we demonstrate the analytical power of this method by using the feeding curves of
worms treated with RNAi for QTL mapping. We tested a panel of 12 genes (tag-214, mel-26,
rab-5, pos-1, par-6, gld-1, let-502, lin-31, par-1, smo-1 and mpk-1) on a recombinant inbred
population derived from a cross between Bristol and a natural isolate from Hawaii. Hawaii has
lost RNAi sensitivity in the germline. We confirmed that polymorphisms in ppw-1 contribute to
this loss of RNAi sensitivity as for many RNAi treatments the QTLs are found at the ppw-1
locus, but moreover we found that other loci are also likely to be important.
In summary, we have established a fast method that improves the throughput of RNAi in liquid,
that generates quantitative data, that is easy to implement in most laboratories, and importantly
that enables QTL mapping using RNAi.
Funding: the EU 7th Framework Programme under the Research Project PANACEA, contract nr
222936.
| Original language | English |
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| Title of host publication | Abstracts of papers presented at the Evolution of Caenorhabditis and Other Nematodes, Cold Spring Harbor Laboratory, New York, USA, 3-6 April 2012 |
| Pages | 46-46 |
| Publication status | Published - 2012 |
| Event | Evolution of Caenorhabditis and Other Nematodes, Cold Spring Harbor Laboratory, New York, USA - Duration: 3 Apr 2012 → 6 Apr 2012 |
Conference
| Conference | Evolution of Caenorhabditis and Other Nematodes, Cold Spring Harbor Laboratory, New York, USA |
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| Period | 3/04/12 → 6/04/12 |