SBIR Phase I: High-Throughput Multi-Analyte ChIP Assay Development

SBIR 第一阶段：高通量多分析物 ChIP 检测开发

• 批准号: 1142432
• 负责人: MaryAnne Jelinek
• 金额: $ 15万
• 依托单位: Active Motif, Inc.
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1142432&HistoricalAwards=false
• 关键词: SBIR; Phase; Throughput; Multi; Analyte

This Small Business Innovation Research (SBIR) Phase I project proposes to develop a high-throughput, multi-analyte chromatin immunoprecipitation (ChIP) system. ChIP is a widely used technique among life science and biomedical researchers seeking map the specific DNA sequences that are bound by a DNA binding protein. Its use ranges from single gene analysis through to genome-wide applications that use next generation sequencing (NGS) technologies as readout. As envisioned, the proposed technology will transform the lengthy and cumbersome multi-day ChIP into a high throughput compatible single-day experiment. The technology will result in the insertion of oligonucleotides containing NGS platform-compatible tags and "bar-code" sequences into DNA fragments associated with the immunoprecipitating antibodies. Each antibody will be associated with a unique barcode that can also be used for direct DNA sequencing or PCR analysis. Feasibility of this approach will be achieved by systematic identification of optimal conditions for insertion of the bar-code containing oligonucleotides into chromatin, and then, with antibody-coupled oligonucleotides. Final validation will be performed at the genome scale using NGS to compare the genomic representation of the library produced by the novel method with that of traditional ChIP. The broader impact/commercial potential of this project is far reaching. The life sciences research tools market is currently estimated at $42 billion, with the epigenetic sector experiencing high growth fueled by researchers purchasing commercial epigenetic products rather than spending the time and effort to develop them in-house, and by advances in NGS, which has accelerated genome-wide epigenetic analyses. Successful development of the high-throughput, multi-analyte ChIP will have significant impact scientifically and commercially in the life sciences and biomedical research arenas. In less than five years post-launch, it is projected to replace traditional ChIP, which represents 20-25% of the epigenetic research tools market estimated at $175-245M in 2010. The development of this method will open epigenetic analysis to virtually all researchers by eliminating technical barriers, and by significantly reducing sample size requirements associated with traditional ChIP, including a potential for single cell analysis. Development of this technology will spur the creation of additional novel technologies such as homogeneous ChIP for high throughput screening, multi-analyte ChIP, and open the door for environmental, nutrition, and toxicology disciplines to study the epigenetic profiles of any eukaryotic organism on a genome wide scale.

这个小企业创新研究（SBIR）第一阶段项目提出开发一个高通量，多分析物染色质免疫沉淀（ChIP）系统。ChIP是生命科学和生物医学研究人员广泛使用的技术，旨在绘制由DNA结合蛋白结合的特定DNA序列。其用途范围从单基因分析到使用下一代测序（NGS）技术作为读出的全基因组应用。正如所设想的那样，所提出的技术将把冗长而繁琐的多日ChIP转化为高通量兼容的单日实验。该技术将导致将含有NGS平台兼容标签和“条形码”序列的寡核苷酸插入与免疫沉淀抗体相关的DNA片段中。每种抗体将与也可用于直接DNA测序或PCR分析的独特条形码相关联。该方法的可行性将通过系统鉴定将含有条形码的寡核苷酸插入染色质中的最佳条件，然后用抗体偶联的寡核苷酸来实现。将使用NGS在基因组规模上进行最终验证，以比较通过新方法产生的文库的基因组表示与传统ChIP的基因组表示。该项目的广泛影响/商业潜力是深远的。生命科学研究工具市场目前估计为420亿美元，表观遗传领域经历了高增长，这是由于研究人员购买商业表观遗传产品，而不是花费时间和精力在内部开发它们，以及NGS的进步，这加速了全基因组表观遗传分析。高通量、多分析物ChIP的成功开发将在生命科学和生物医学研究领域的科学和商业上产生重大影响。 在推出后不到五年的时间里，它预计将取代传统的ChIP，后者占2010年表观遗传研究工具市场的20-25%，估计价值1.75 - 2.45亿美元。这种方法的开发将通过消除技术障碍，并通过显着减少与传统ChIP相关的样本量要求（包括单细胞分析的潜力），向几乎所有研究人员开放表观遗传分析。这项技术的发展将刺激创造更多的新技术，如用于高通量筛选的均质ChIP，多分析物ChIP，并为环境，营养和毒理学学科打开大门，以研究任何真核生物在全基因组范围内的表观遗传特征。