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Genome-wide pooled CRISPR screen in arthropod cells

Published on April 30, 2024   27 min

A selection of talks on Genetics & Epigenetics

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0:00
My name is Norbert Perrimon from Harvard Medical School and the Howard Hughes Medical Institute. Today, I'm going to tell you about genome-wide pooled CRISPR screen in arthropod cells. My lab mostly works in Drosophila, but I'm going to describe the technology that we established first in flies, and that now we're extending to mosquitoes, and also tick. As you know, Drosophila is a modern system of functional genomics and it's mostly for in vivo studies, but today I'm going to be describing the applications in Drosophila cells.
0:35
Why do we want to perform the functional genomic screens in Drosophila cells? Flies are very well-known for the conservation of many different signaling mechanisms and most of the signaling pathways that we know about are conserved in flies. One of the interests about the forming screens in Drosophila are that there is a lot less redundancy in the fly genome compared to mammalian systems but in addition, there are a number of different processes, which can only be studied in the fly cells or the arthropod cells. There are a number of signaling pathways like ecdysone signaling, that we've described briefly in my talk, are very specific to insects. There are also a number of different pathogens that are insect-specific. Again, a number of different toxins and viruses can only be analyzed in the insect set.
1:26
With regard to functional genomics methods, most of the approaches we are going to be discussing about in the cell lines are either RNAi or CRISPR. So, with RNAi, the introduction of double-stranded RNAs, either long double-stranded RNAs (dsRNAs) or short-hairpin RNAs (shRNAs) are going to perturb the RNA, the endogenous RNA, so this can be used to generate partial loss of function perturbations. With CRISPR on the other hand, the CRISPR is a gene-editing technology where here in this case we are going to generate double-strand breaks which are going to induce mutations in the targeted genes.

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Genome-wide pooled CRISPR screen in arthropod cells

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