Gene expression patterns in different cell types can be compared
three basic types that differ in their immobilised (target) DNA
changes in gene expression level (transcriptomic)
expression chips
immobilised DNA is cDNA derived from RNA
can be used to study ......
diseased vs healthy gene expression
development changes in gene expression
changes to an altered metabolic environment
genomic gains and losses (genomic)
DNA repair genes
comparative genomic hybridisation
immobilised DNA are large pieces of genomic
DNA with a known chromosomal location
fluorescence reflects copy number
mutations in DNA
detects mutations and polymorphisms
immobilised DNA is a single gene with many variants
High-throughput screening
method used for scientific experimentation such as drug discovery
allows a researcher to conduct millions of biochemical, genetic and pharmacological testing
can identify active compounds, antibodies or genes which modulate a particular biomolecular pathway
results provide starting points for drug design and for understanding the interaction or role of a particular biochemical process in biology
target validation
makes sure the role of the inhibitor in the
disease is thoroughly known and the regulation
of the target cell has the expected affect
allows the identification of compounds
whose inhibition results in cell apoptosis or
the inhibition of cell proliferation
primary screening
large scale primary screening in living cells to select
the best hits among thousand of compounds
secondary screening
panel of assays for focused secondary screening in cell-based models of human diseases or in primary cell cultures
lead optimisation
identify compounds suitable for testing in a clinical setting
GFP-Tag
GFP = Green Fluorescent Protein
originally isolated from the jellyfish Aequorea victoria
30 members of GFP-like
family from Anthozoans
naturally occurring 27 kD protein that fluoresces green when exposed to blue light
useful in fluorescence microscopy techniques
Anticytokines
cytokines are important components
of the immune system
imbalance of the cytokine process (production,
receptor expression and dysregulation)
contributes to many pathological disorders
there are pathways from gene
expression to protein effect
there are also many potential points of intervention
is being analysed for possible therapeutic uses
Image analysis
designed to provide continuous, time‐lapse images of live cells from within your own standard cell‐culture incubator
can produce a graph showing kinetic proliferation curves (proliferation rates against time)
FACS
FACS = Fluorescence Activation Cell Sorting
flow cytometryis a technique for counting and examining
microscopic particles, such as cells and chromosomes, by
suspending them in a stream of fluid and passing them by an
electronic detection apparatus
allows simultaneous multiparametric analysis of the
physical and/or chemical characteristics of up to
thousands of particles per second