Development of a method to multiplex ChIP-SEQ and ChIP-chip experiments

Award Information
Agency:
Department of Health and Human Services
Branch
n/a
Amount:
$96,738.00
Award Year:
2010
Program:
SBIR
Phase:
Phase I
Contract:
1R43HG005282-01
Agency Tracking Number:
HG005282
Solicitation Year:
2010
Solicitation Topic Code:
NHGRI
Solicitation Number:
PHS2010-2
Small Business Information
AFFOMIX, INC.
AFFOMIX, INC., 688 East Main Street, Branford, CT, 06405
Hubzone Owned:
N
Socially and Economically Disadvantaged:
N
Woman Owned:
N
Duns:
623623803
Principal Investigator:
MICHAEL WEINER
(203) 458-2844
MICHAEL@MWEINER.COM
Business Contact:
(203) 458-2844
mweiner@affomix.com
Research Institution:
n/a
Abstract
DESCRIPTION (provided by applicant): The immediate objective of our research is to generate a method that will enable researchers to multiplex Chromatin ImmunoPreciptation-Sequencing (ChIP-Seq) analysis in a single Next generation DNA sequencing run. In the to-be-developed method: i) a set of antibodies directed against specific DNA-binding proteins are uniquely bar-coded with a DNA 'ZipCode;' ii) covalent cross-links between DNA-binding proteins and chromosomal DNA are formed by treating cells with formaldehyde; iii) the set of DNA-barcoded antibodies specific to the proteins of interest are used to selectively coimmunoprecipitate the protein-bound DNA fragments that were covalently cross-linked; iv) excess antibodies and chromosomal DNAs are removed by washing; v) the enriched protein-bound chromosomal DNA is ligated to the antibody-attached ZipCode DNA using T4 DNA ligase, and; vi) the immunoprecipitated protein-DNA links are reversed and the recovered DNA is assayed using Next-generation sequencers to determine both the chromosomal DNA sequence bound by the protein and the antibody-identifying DNA ZipCode. PUBLIC HEALTH RELEVANCE: Researchers will benefit from an increased understanding of the function of human proteins. Methods are needed that can facilitate this understanding. The immediate objective of our research is to generate a method that will enable researchers to multiplex the method known as Chromatin ImmunoPreciptation-Sequencing (ChIP-Seq) analysis, in a single DNA sequencing run. We anticipate using our high-throughput antibody-discovery pipeline for producing recombinant antibodies as a means to generate the reagents needed to make this multiplexed method possible.

* information listed above is at the time of submission.

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