Histone modifications carry rich information of cellular memory and gene regulatory mechanisms. Single cell analysis of histone modification in conjunction with transcriptome could recover this critical layer of cell identity and help to dissect the cellular and molecular composition of complex tissues such as the brain. We recently developed an ultra-high throughput single cell multi-omics assay, Paired-Tag, that can jointly map transcriptome and histone modifications from up to a million single cells in parallel. In order to translate this novel technology to the marketplace and to advance the goals of the BRAIN initiative, we propose to optimize and further develop Paired-Tag to enable its broad use in the research community. Specifically, we will develop Paired-Tag kits to allow for general accessibility of the method in molecular biology laboratories. We will then carry out extensive field tests of the Paired-Tag kit in the laboratories of current BRAIN initiative investigators, and further improve it based on the feedbacks. Finally, we will adapt the Paired-Tag procedure to long-read sequencing technologies to enable high throughput detection of splicing isoforms in brain cells at single cell resolution. If successful, the proposed research would provide researchers with a powerful new tool for single cell transcriptomics and epigenomics analysis.
Public Health Relevance Statement: We proposed to translate a novel technology supported by Brain Research through Advancing Innovative Neurotechnologies (BRAIN)-initiative to the marketplace, to help investigators to better map the inner workings of brain cells at single cell resolution and dissect the organization and function of the neural circuits.
Project Terms: Adoption; Adult; 21+ years old; Adult Human; adulthood; Alternative Splicing; Alternate Splicing; Alternative RNA Splicing; Antibodies; Bar Codes; barcode; Biological Assay; Assay; Bioassay; Biologic Assays; Brain; Brain Nervous System; Encephalon; Complementary DNA; cDNA; Cells; Cell Body; Chromatin; Communities; DNA; Deoxyribonucleic Acid; Equipment; Feedback; Genes; Regulator Genes; Transcriptional Regulatory Elements; regulatory gene; trans acting element; Goals; Hippocampus (Brain); Ammon Horn; Cornu Ammonis; Hippocampus; hippocampal; Human; Modern Man; Institutes; Joints; Laboratories; Lead; Pb element; heavy metal Pb; heavy metal lead; Ligation; Closure by Ligation; Manuals; Maps; Memory; Methods; Molecular Biology; DNA Molecular Biology; Mus; Mice; Mice Mammals; Murine; Glia; Glial Cells; Kolliker's reticulum; Neuroglial Cells; Non-neuronal cell; Nonneuronal cell; nerve cement; Neuroglia; Nerve Cells; Nerve Unit; Neural Cell; Neurocyte; neuronal; Neurons; Quality Control; Reagent; Research; Research Institute; Investigators; Researchers; Research Personnel; Reverse Transcription; Splicing; RNA Splicing; Technology; Testing; Tissues; Body Tissues; Translating; Translations; Work; falls; Tube; base; improved; Procedures; Area; Phase; Link; Ensure; Individual; Licensing; Funding; Cell Lineage; tool; catalog; Catalogs; programs; cognitive function; Adopted; Complex; Protocol; Protocols documentation; cell type; brain tissue; interest; brain cell; field based data; field learning; field test; field study; Performance; Isoforms; Protein Isoforms; single cell analysis; Structure; novel technologies; new technology; Modality; neural function; Neurophysiology - biologic function; neural circuitry; neurocircuitry; synaptic circuit; synaptic circuitry; neural circuit; DNA bank; DNA Library; histone modification; Address; Length; Detection; Resolution; Small Business Technology Transfer Research; STTR; Transcript; Molecular; Process; Development; developmental; epigenomics; anticancer research; anti-cancer research; cancer research; design; designing; transcriptomics; brain research; operation; epigenome; BRAIN initiative; Brain Research through Advancing Innovative Neurotechnologies initiative; multiple omics; multiomics; transcriptome; global gene expression; global transcription profile; single cell technology; experimental study; experiment; experimental research; innovative neurotechnologies; DNA sequencing; DNA seq; DNAseq; neural network
