B2B - Target Validation

Question 1

Target Validation Assays (5)

Answer 1

1. Biochemical and cell based
2. Yeast (mutant phenotypes)
3. Appropriate animal models
4. Knockout and knockin
5. Zebra fish-transgenic. Established physiology for disease

Question 2

Target Validation questions to ask

Answer 2

1. Expression in disease organs/tissues
2. Can the disease phenotype be alleviated using RNAi/gene knockout
3. Is disease phenotype induced by knockin of target
4. Is the target druggable

Question 3

RNA technology

Answer 3

1. dsRNA specific to gene of interest for silencing is introduced activating RNAi pathway
2. Dicer cleaves dsRNA to short double stranded fragments
3. These are separated into single strands (siRNA) and introduced into RISC (RNA-induced silencing complex)
4. siRNAs basepair to target mRNA inducing mRNA cleavage preventing its function as translation template

Question 4

mABs advantages over small molecules

Answer 4

good as they interact with larger surface of target. Allows for discrimination between highly similar targets. reducing chance of off-target small molecule toxicity

Question 5

e.g. receptors

Answer 5

P2X7 receptor and pain
HER2 (EGFR) and cancer
S1P1 and MS
cAMP, PDE and COPD - oxidative stress and CD8 cell T 
cell. Roflimulast on PDE4

Question 6

Chemical genomics as a strategy for target validation

Answer 6

study of gene responses to chemical compounds.
Chemical libraries with high-information content cellular assays and using data-analysis tools.
Evaluate cellular function before committing to the next part of drug development