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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.
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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.
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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.
