Detection of Methylation and Translocation Events by Novel Genomic Technologies

Detection of Methylation and Translocation Events by Novel Sequencing Technology ABSTRACTThe rapid development of DNA sequencing technologies advanced our understanding of the molecular basis of cancerHowevercancer genomics involves very complex molecular rearrangementssuch as translocationsrequiring the investigation of long DNA molecules with single nucleotide discriminationCurrent Next Generation SequencingNGStechnologies cannot address such performance requirements and face challenging limitationsThe long term goal of this proposal is to develop a rapid and highly accurate DNA sequencing system that has the potential to fully control the speed and orientation at which the DNA passes through an electronic detectorsolid stateboth in forwardreadingand reverseproof readdirectionsIn contrast with other nanopore technologiesour technology decouples the feeding step from the reading stepenabling to sequence full genomes starting with minute quantity of materialCombinedthe characteristics have the potential to provide our sequencing technology with the capability to achieve single nucleotide discrimination with an accuracy ofor higher using long DNA moleculeskbSuch a system is anticipated to be capable of accurately reading such long fragments of DNA starting from minute quantities of material without the need of amplificationThis would provide both practitioners and cancer researchers with a much better tool than the current state of the art sequencing technologiesIndeed we project that our technology will enable the collection of information of genetic and epigenetic markers of cancer both in expressed and non expressed areas of the genome while reaching time and cost performance metrics currently unachieved on the marketThe goal of the current PhaseRproposal is to demonstrate that by incorporating our proprietary technology into a sequencing systemwe will be able to detect important molecular changes that are associated with cancer initiation and proliferationto include long range methylations and translocations as well as chromoplexySuch detection will be rendered possible due to the ability of our system to fully control the translocation rate and linearization of the DNA while passing through the sequencing system detectorIn Phase IIRwill focus on the incorporation of an embedded solid stateelectronicnanoelectrode detection component to the initial system developed in phase I for single base nucleotide readingProvided that targeted milestones are achievedcommercial development of a prototype instrument along with associated consumable chips and reagents will be initiated immediately following the completion of phase IIWe project that our technology has the potential to sequence a full human genome in less than one hour at the cost of $from biopsy materialthus reaching performance metrics currently unavailable on the current marketand providing cancer researchers and medical practitioners with an invaluable affordable tool for the elucidation of molecular mechanisms of cancer and the detection of specific molecular changes which may improve early clinical detection of the disease Narrative The potential of high accuracy sequencing of long DNA moleculesbpsfor cancer research and clinical diagnostics applications is well recognized by the scientific and medical communityThe ability to map methylation and long range translocation events with a high degree of accuracy will significantly contribute to advancements in the field of cancer diagnostics especially if such sequencing can be performed with single base resolutionat a cost of less than $within minutesTo datethe best technologies for genome mapping sequencing have failed to satisfy the demand for accuracyspeedcostand long DNA fragments which is essential for translocation event detection