E.R. Gauthier - Grantsmanship 101 – March 2011
Grantsmanship 101 – Experimental design.
1) Ground your proposal in reality: pipedreams never get funded.
a. Clear hypothesis
b. Preliminary data always help
c. Clearly defined aims (i.e. research objectives)
d. Appropriate experimental design
e. Appropriate methods for data analysis (e.g. stats)
f. Highlight pitfalls and means to counter them
2) Appropriate research model
a. Know your favorite cell line/animal (e.g. mutated genes, etc)
b. Justify why you selected this cell line/ that animal.
c. Beware of strain-specific effects: include more than one cell type/animal strain
d. Stable transfection? Analyze at least 3 clones per construct to control for clonerelated effects.
e. Transient transfection: how will you transfect the cells? What is the transfection
efficiency of your favorite cell line? How will you detect the transfected cells
over the background of untransfected cells?
f. Beware of methods based on overexpression (inducible expression can afford
much better control over the expression levels  more “physiological”).
g. Beware of mutants (e.g. cancer cell lines vs real cells)!
3) Availability of reagents
a. Inhibitors: beware of poor quality  carefully select your provider of inhibitors.
b. Antibodies: if unavailable, propose viable alternative, e.g. use an epitope-tagged
version of your favorite protein. AVOID having to make the antibody. If you
really, but REALLY need to make it, contract the work to a specialized company
(unless you have documented experience in antibody production).
c. Constructs: have them at hand before you send the grant.
d. Collaborators: if you rely on someone else to provide you with reagents, make
sure they can do so for the entire duration of your funding cycle (provide support
letter with the grant if possible/allowed).
4) Specificity of reagents
a. Inhibitors: very, VERY few inhibitors are really, REALLY specific. Always
propose an alternate means to block your favorite pathway/enzyme that will not
make use of the inhibitor (e.g. RNAi, dominant negative constructs)
b. Antibodies: are they REALLY specific to your favorite protein? Can they detect
different isoforms of the same protein? Can they cross-react with multiple
proteins of the same family? Using a competing peptide corresponding to the
epitope can help convince the reader of the specificity of your assay.
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E.R. Gauthier - Grantsmanship 101 – March 2011
c. RNA interference: currently the hottest field with the fewest controls! Always
control for off-target effects and for general vs specific down-regulation of gene
expression.
5) Feasibility:
a. Do you have the expertise?
b. Do you have the facilities?
c. Can graduate students do the work, or will you rely on PDFs/research associates?
d. Can the proposed research realistically be done during the proposed funding
cycle?
6) Pitfalls:
a. Is there more than one interpretation possible for your data?
b. What if your hypothesis is wrong? What will you do then? Provide a back-up
plan.
c. Have other people obtained results that support/counter your hypothesis (e.g. in a
completely different animal)?
7) “Discovery-driven” research
a. Beware of fishing expeditions.
b. Make it a clearly defined part of an aim/the proposal (grant selection committees
usually do not like fishing expeditions  give them the opportunity to fund you
for the other stuff you are proposing if they don’t like your fishing trip).
c. Know what you’re looking for (the worst thing you can do is say “I’ll get
candidate proteins/genes and I’ll go from there”).
d. Provide a stringent strategy to select candidates for further validation (in other
words, how will you determine whether you caught a nice trout vs an old shoe).
e. Provide a clear candidate validation scheme.
f. Beware of the limitations of your screen (rate of false positives, sensitivity [will
you miss low abundance targets?], too many ESTs).
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E.R. Gauthier - Grantsmanship 101 – March 2011
HYPOTHESIS
Caspase 8 is involved in
the alcohol-induced
death of my brain cells
Measure
Caspase 8
activity
Sub-cellular
localization
of Casp8
Status of
known
Casp8
targets
Block Casp8
Chemical
inhibitors
Casp8-/cells/mice
RNAi
Dominant
negative
cDNAs
3
Overexpress
Casp8
Inducible
Wild
type
Constitutive
Constitutively
active