Biology Reference
In-Depth Information
CHAPTER EIGHT
Exploiting Drosophila Genetics
to Understand MicroRNA Function
and Regulation
Qi Dai, 1 Peter Smibert, 1 and Eric C. Lai
Contents
1. Introduction
202
204
2. Generation of miRNA Mutants
2.1. Transposon-induced miRNA mutants from forward genetic
screening
204
208
2.2. Deletion of miRNA loci by imprecise excision of P elements
2.3. Other transposons
209
210
2.4. miRNA alleles induced by homologous recombination
2.5. On the importance of rescuing mutant phenotypes
212
213
3. The Gal4-UAS System for Transgene Activation
3.1. Misexpression of miRNAs from genomic EP insertions
214
216
3.2. Directed analysis of UAS-miRNA transgenes
217
3.3. miRNA sponges
218
4. Genetic Interactions and Epistasis Tests
218
4.1. Double mutant analysis to uncover redundant functions
4.2. Epistasis analysis
219
220
4.3. Rescue of miRNA mutants by target heterozygosity
4.4. Dominant modifier screens for miRNA interactors
222
223
5. Detecting miRNA Expression and Activity In Vivo
5.1. miRNA promoters “identified” by P elements
224
224
5.2. miRNA enhancer:reporter transgenes
5.3. Genetically encoded sensors of miRNA activity
226
228
6. Conclusions and Future Prospects
Acknowledgments
230
References
230
Department of Developmental Biology, Sloan-Kettering Institute, New York, USA
1 These authors contributed equally.
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