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Reagent Development for Small Cell Number ChIC-seq


Fast-Track proposals will not be accepted.
Number of anticipated awards: 1 Budget (total costs):
Phase I: $150,000 for 1 year
It is strongly suggested that proposals adhere to the above budget amounts and project periods. Proposals with budgets exceeding the above amounts and project periods may not be funded.
This solicitation is for the development of conjugates between specific antibodies or protein A with microccocal nuclease (MNase) to be used for genome-wide epigenetic mapping. They will be used to identify genome-wide epigenetic changes during normal development and pathological conditions, requiring only a few hundred primary or patient cells.
Project Goals
The project goal is to develop reagents that can be used for mapping genome-wide epigenetic changes during normal development and disease process in rare primary and patient cells. Because conjugating proteins could result in inactivation of the proteins, it will be important to achieve efficient conjugation between antibodies and MNase while reserving the activities of both.
Phase I Activities and Expected Deliverables
Specific deliverables are:
 Conjugates between 10 different histone modification antibodies and MNase; one milligram specific antibody per
histone modification (antibodies include H3K4me1, H3K4me2, H3K4me3, H3K27ac, H3K27me3, H2A.Z, H3K9ac,
H3K9me3, H3K36me3, histone H3).
 Conjugates between 40 transcription factor antibodies and Mnase; one milligram specific antibody per transcription
factor (antibodies for transcription factors include RNA Pol II, Brd4, BRG1, GATA3, Eomes, T-bet, ETS1, RORg,

and other general and sequence-specific factors that will be decided by the Contracting officer Representative (COR) on the resultant contract.  Conjugates between Protein A and MNase (10 milligrams)
It is critical that the developer provide evidence to show that the antibodies in the conjugates are still specific and as active as non-conjugated antibody using Western and ChIP assays. It is critical that the MNase in the conjugates is still as active as non-conjugated MNase using chromatin digestion assay. It is critical that the free individual proteins (un-conjugated) in the final products are less than 5% of all protein components.

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