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Printable Handouts
Navigable Slide Index
- Introduction
- Why do screens in fly cells?
- RNAi and CRISPR based screening
- Timeline of genome-wide screening
- Arrayed genome-wide RNAi screening
- Arrayed RNAi screens at the DRSC
- Timeline of genome-wide screening (recently)
- Pooled CRISPR screening
- RMCE-CRISPR
- Pooled sgRNA delivery by phiC31 integration
- CRISPR knockout genome-wide library
- Essential genes are enriched for conservation
- Drug proliferation screens in Drosophila cells (1)
- Drug proliferation screens in Drosophila cells (2)
- FK506-bp2/FKBP12 mediates rapamycin inhibition of mTOR
- Ecdysone enters cells & causes proliferative arrest
- A CRISPR KO screen for reduced ecdysone response
- Ecdysone importer (EcI) is necessary
- EcI identification argues for a new view of regulation
- Tripartite pore-forming complexes toxins production
- Tc toxins causes dose-dependent cell enlargement
- Mode of entry of P. Luminescence Tc toxin
- P. luminescens e recognizes visgun (Vsg)
- Vsg knockout blocks Tc-induced effects
- Vsg is required at low pTc concentration
- ESCRT0 & HOPS genes are lost in Vsg KO screen
- Many hits are in the N-linked glycosylation complex
- MGAT1/2 were top hits in a HeLa cell CRISPR screen for pTc
- Tc toxins are produced by insect and human-pathogenic bacteria
- A screen using M. morganii Tc toxin
- CRISPR screens in mosquito cells
- Identification and evaluation of pol II promoters
- Pooled screen in Anopheles cells
- Pooled screens in Anopheles
- Pooled screening in tick (I. scapularis) cells
- CRISPR screens in arthropods (1)
- Pooled CRISPR activation screening
- Pooled CRISPRa screening
- Genome-wide CRISPR activation screen for rapamycin resistance genes
- InR-Akt-mTOR activation
- CRISPR screens in arthropods (2)
Topics Covered
- RNAi based screening
- CRISPR based screening
- Genome-wide screening
- Drosophila S2R+ cells
- Ecdysone
- Tripartite pore-forming complexes (Tc) toxins
- P. luminescence
- Visgun (Vsg)
- M. morganii
- Rapamycin resistance
Links
Series:
Categories:
External Links
Talk Citation
Perrimon, N. (2024, April 30). Genome-wide pooled CRISPR screen in arthropod cells [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved December 3, 2024, from https://doi.org/10.69645/QCKW8305.Export Citation (RIS)
Publication History
Financial Disclosures
- There are no financial/commercial matters to disclose.
A selection of talks on Genetics & Epigenetics
Transcript
Please wait while the transcript is being prepared...
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.