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

Award Information
Agency: Department of Health and Human Services
Branch: N/A
Contract: 1R43HG005282-01
Agency Tracking Number: HG005282
Amount: $96,738.00
Phase: Phase I
Program: SBIR
Awards Year: 2010
Solitcitation Year: 2010
Solitcitation Topic Code: NHGRI
Solitcitation Number: PHS2010-2
Small Business Information
AFFOMIX, INC., 688 East Main Street, Branford, CT, 06405
Duns: 623623803
Hubzone Owned: N
Woman Owned: N
Socially and Economically Disadvantaged: N
Principal Investigator
 (203) 458-2844
Business Contact
Phone: (203) 458-2844
Research Institution
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.

Agency Micro-sites

US Flag An Official Website of the United States Government