Genome-Wide High Content Screens of Cell Function and Behaviour
Jeremy Simpson, Professor, University College Dublin
In cells, molecules are transported
between membrane-bounded compartments by the process of membrane traffic. Due
to the exquisite morphology and distribution of organelles in mammalian cells,
imaging approaches are highly suited to the study of these transport pathways.
Broadly, membrane traffic pathways con be categorised as secretory (endoplasmic
reticulum to cell surface), endocytic (cell surface to endosomal-lysosomal
system) and retrograde (endosome to endoplasmic reticulum). We are using a
combination of genome-wide RNA interference and automated high content
screening microscopy to systematically dissect each of these pathways, and
quantify the relevance of all genes to them. Of particular interest is the
identification of regulatory networks that span all three pathways. In
addition, we are using the information from these screens to understand the
mechanism by which synthetic nanoparticles interact with cells and then enter
them. This information is fundamental to our design of next generation drug delivery
vectors.
Results
from the screens indicate a surprising level of pathway complexity, and links
to families of proteins never previously anticipated. The work also highlights
the power of quantitative analysis of organelle morphology, particularly using
image texture features, as a tool to identify regulatory machinery. This
analyses will ultimately provide a complete systems-level view of how cells are
organised, and will inform how they interact with nanosurfaces and
next-generation biomedical devices.
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