Genome-wide quantification of rare mutation by Bottleneck Sequencing

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Unmet Need
The underlying mechanisms of aging, carcinogenesis, and neurodegeneration involve the buildup of stochastic somatic cell mutations. Understanding the temporal and spatial properties of these mutations can lead to a more mechanistic understanding of human disease progression and the design of genetic-level therapeutics.
No simple method of high-throughput genetic screening of somatic point mutations currently exists with sufficient sensitivity to identify these rare mutations. Most modern techniques use single cell genomic sequencing, which can detect these rare mutations genome-wide but cannot identify such variations in a single cell.
Technology Overview
The Bottleneck Sequencing System (BotSeqS) uses consensus sequencing with molecular barcodes to identify rare somatic point mutations in single cells with high sensitivity. The BotSeqS method involves diluting a sequencing library prior to applying standard PCR amplification techniques. This dilution creates a bottleneck during library sampling, allowing optimal random processing of both DNA strands with efficient sequencing. Identification of rare mutations on both DNA strands provides increased detection specificity. This algorithm allows these uncommon mutations to be unmasked among the abundance of wild-type sequences contained in most common libraries.
Stage of Development
The inventors have shown the application of the BotSeqS technology to identify and quantify the existence of rare somatic point mutations in genomic sequences from healthy human tissues. This initial study showed BotSeqS accurately measured age and tissue-dependent accumulations of rare mutations. Additionally, the inventors provided evidence that BotSeqS can differentiate the mutation patterns in both mitochondrial and nuclear genomes and correlate the mutation spectra with common cancerous cells.
Hoang ML et al., Proc Natl Acad Sci U S A. 2016 Aug 30;113(35):9846-51
Patent Information:
Title App Type Country Serial No. Patent No. File Date Issued Date Expire Date Patent Status
BOTTLENECK SEQUENCING PCT: Patent Cooperation Treaty United States 16/073,622 7/27/2018     Pending
For Information, Contact:
Jeanine Pennington
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