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- Introduction
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1. Drosophila genetics - the first 25 years
- Prof. Dan Lindsley
- Establishment of the Primary Body Axes
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2. Homeotic genes in Drosophila's bithorax complex - The legacy of Ed Lewis
- Prof. Francois Karch
- Cell Type Specification and Organ Systems
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4. From germ cell specification to gonad formation
- Prof. Ruth Lehmann
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5. Drosophila stem cells
- Prof. Michael Buszczak
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6. Legacy of drosophila genetics: female germline stem cells
- Prof. Michael Buszczak
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7. Intestinal stem cell-mediated repair in Drosophila 1
- Prof. Tony Ip
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8. Intestinal stem cell-mediated repair in Drosophila 2
- Prof. Tony Ip
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10. Axon guidance in Drosophila
- Prof. John Thomas
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11. Development and physiology of the heart
- Prof. Rolf Bodmer
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12. Identification of host defenses in the Drosophila gut using genome-scale RNAi
- Prof. Dominique Ferrandon
- Genome Organization and Function
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13. The genetic analysis of meiosis in Drosophila melanogaster females
- Prof. R. Scott Hawley
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15. Dorsal-ventral patterning of the Drosophila embryo
- Prof. Mike Levine
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17. Genome-wide pooled CRISPR screen in arthropod cells
- Prof. Norbert Perrimon
- Behavior
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19. Genetics of chemosensory transduction: taste and smell
- Dr. Leslie Vosshall
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20. Cracking the case of circadian rhythms by Drosophila genetics
- Prof. Jeffrey C. Hall
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21. Sleep in Drosophila
- Dr. Ralph Greenspan
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23. Drosophila as a model for drug addiction
- Prof. Ulrike Heberlein
- Mechanism of Human Disease
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24. Cross-genomic analysis of human disease genes
- Prof. Ethan Bier
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25. Human neurodegenerative disease: insights from Drosophila genetics
- Prof. Nancy Bonini
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26. Metastasis of Drosophila tumors
- Prof. Allen Shearn
- Evolution of Adaptive Novelties
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28. The evolution of morphological novelty
- Prof. Nipam Patel
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29. The genetic architecture of complex traits: lessons from Drosophila
- Prof. Trudy Mackay
- Archived Lectures *These may not cover the latest advances in the field
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30. Using gene expression information to provide insights into patterning and differentiation
- Prof. Angelike Stathopoulos
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31. Regulation of gastrulation in Drosophila
- Prof. Dr. Maria Leptin
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32. microRNA function in stem cells
- Prof. Hannele Ruohola-Baker
Printable Handouts
Navigable Slide Index
- Introduction
- Functional genomics
- Functional analysis in Drosophila
- RNA-interference (RNAi)
- Genetics by RNAi in Drosophila cells
- Genetic screens by RNAi
- Overview of cell-based RNAi screens
- Libraries for Drosophila RNAi screens
- Generation of Drosophila dsRNA library
- Considerations in library design
- Cell-based assays for RNAi screens
- Decisions in design of cell-based assays
- Detection methods for cellular assays
- Analysis of RNAi screens
- Systematic analysis of high-throughput data
- Processing of high-throughput assays
- Normalization and hit identification
- Challenges in quantitative image analysis
- Issues and insights from screen results
- Applications for RNAi screens: systematic
- Assay for cell fitness
- Genome-scale RNAi analysis of cell 'fitness'
- Comprehensive identification of essential genes
- Assay for cell morphology
- Image acquisition
- Genes identified by visual RNAi phenotypes
- Spatial affects on F-actin organization
- Shared defects - shared pathways
- Applications for RNAi screens: comparative
- Sets of genes affect specific morphology
- Applications for RNAi screens: reiterative
- RNAi screen for modifiers of dsRNA phenotype
- Outlook for RNAi screens
- Acknowledgements
- Resources for conducting RNAi screens
- Resources for RNAi screen data
Topics Covered
- Functional analysis in Drosophila
- RNAi
- Genetic screens by RNAi
- RNAi libraries
- Cell-based assays
- Screen analysis
- Systematic analysis of high-throughput data
- Normalization and hit identification
- Challenges in quantitative image analysis
- Issues and insights from screen results
- Applications for RNAi screens
- Assay for cell fitness
- Assay for cell morphology
- Shared defects and shared pathways
- Outlook for RNAi screens
Links
Series:
Categories:
Talk Citation
Kiger, A. (2018, May 31). Genome scale analysis of cellular processes using RNAi [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved October 8, 2024, from https://doi.org/10.69645/HGZO4603.Export Citation (RIS)
Publication History
Financial Disclosures
- Dr. Amy Kiger has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.
A selection of talks on Methods
Transcript
Please wait while the transcript is being prepared...
0:00
Hello, I'm Amy Kiger from the University
of California in San Diego.
And I'll be talking to you today about
genome scale analysis of cellular
processes using RNAi.
0:12
So I'll be talking primarily about a new
methodology that has recently come to
Drosophila, and that is to do with
the era of functional genomics.
And this terminology has been
thrown around a lot recently.
But really,
it's classic genetics with a twist in
that it's now sequence informed genetics.
Given the complete genome
sequence that's available.
Simply we can use genome sequence to guide
the systematic perturbations in order to
identify potentially all gene
functions that contribute
to a specific cellular process.
0:46
Drosophila of course has a long history
as a useful model organism for functional
analysis and many genes have been well
characterized now in the organism.
With the advent of complete
genome sequence available,
Drosophila is also now an ideal model for
functional genomic approaches.
And these work conducted in cells and
culture will complement the ongoing
efforts for
in vivo analysis with mutant animals.
So today I'll be talking about
the use of Drosophila cell lines and
information we can learn there
at the cellular level for
gene functions to then make predictions,
as well as provide insights for
ongoing studies in the animal and
during development.
1:28
The discovery of the cellular
process of RNA interference or
RNAi has been advantageous for
the exploitation of this process for
functional genomic approaches
in many organisms.
So what is RNAi?
RNAi occurs when the cells receive double
stranded RNA, serves as a trigger that
then leads to the destruction of cellular
mRNAs that carry homologous sequences.
And although this is a highly
simplified cartoon of the process,
current research has found that
this is highly specific in that
the double stranded RNA is processed
into short interfering RNAs or
siRNAs of 21 base pairs, and that this
21 base pair match to the target RNA is
imperative in order for the destruction
machinery to destroy the target RNA.
So, non specific matches will
not lead to destruction and
only specific matches of
the 21 nucleotides and like.