Single cell RNA sequencing (scRNAseq) provides unprecedented insight into the gene regulatory pathways of living cells in complex tissues. Current single cell technologies can provide high-quality, high resolution analysis of perturbations in cellular populations, but are limited in terms of access, affordability, and scalability of cell input. In order to broadly deploy scRNAseq as a high-throughput, cost effective research and discovery tool, a platform requires several characteristics: sensitivity to detect subtle changes in gene expression as a result of pharmaceutical intervention, scalability in order to efficiently process hundreds to hundreds of thousands of input cells, a protocol capable of processing multiple control and treatment conditions, and efficient multiplexed sequencing capability for cost-effectiveness. Fluent BioSciences has developed Pre-templated Instant Partitions for single cell RNA sequencing (PIPseq) as a novel approach toward scRNAseq that has key advantages over existing methods. PIPseq provides unprecedented accessibility to scRNAseq for applications that are not readily addressable by competing technologies. Eliminating the dependence on complex instrumentation and expensive microfluidic consumables allows even modestly equipped cell biology labs to implement powerful scRNAseq assays. The intrinsic scalability of the PIPseq platform is also particularly useful in applications where many patient samples or treatment conditions might be evaluated. This direct-to-phase II proposal will establish commercial-readiness of PIPseq kits through rigorous internal stability and reproducibility testing, external evaluation and reproducibility testing with multiple genomics core laboratory collaborators, and pre-clinical evaluation of primary breast tissue samples provided by an academic collaborator.
Public Health Relevance Statement: PROJECT NARRATIVE Fluent BioSciences will commercialize Pre-templated Instant Partitions for scRNA sequencing (PIPseq) for sensitive, high-throughput, and scalable single cell genomics. PIPseq enhances accessibility of scRNAseq methods by eliminating complex instrumentation, expensive microfluidic consumables, and efficiently enables large-scale, multisample transcriptomic comparisons across multiple cell populations.
Project Terms: Aging; Bar Codes; barcode; Biological Assay; Assay; Bioassay; Biologic Assays; Biological Sciences; Biologic Sciences; Bioscience; Life Sciences; Breast; Complementary DNA; cDNA; Malignant Neoplasms; Cancers; Malignant Tumor; malignancy; neoplasm/cancer; Capital; Cell Separation; Cell Isolation; Cell Segregation; Cell Separation Technology; cell sorting; Cells; Cell Body; Clinical Research; Clinical Study; cost effectiveness; Estrogens; Therapeutic Estrogen; Experimental Designs; Feedback; Gene Expression; Genes; Regulator Genes; Transcriptional Regulatory Elements; regulatory gene; trans acting element; Human; Modern Man; indexing; Industry; instrumentation; Laboratories; Libraries; Methods; Molecular Biology; DNA Molecular Biology; Patients; Production; Corpus Luteum Hormone; Delta4-pregnene-3,20-dione; Pregn-4-ene-3,20-dione; Pregnenedione; Therapeutic Progesterone; Progesterone; Reagent; Research; Investigators; Researchers; Research Personnel; Technology; Testing; Time; Tissues; Body Tissues; Genetic Transcription; Gene Transcription; RNA Expression; Transcription; Generations; Population Heterogeneity; diverse populations; heterogeneous population; population diversity; Immunology; base; Site; Clinical; Phase; Variant; Variation; Physiological; Physiologic; Evaluation; insight; wasting; Individual; Logistics; Contracting Opportunities; Contracts; tool; Life; cell biology; Cellular biology; Adopted; Complex; Dependence; Protocol; Protocols documentation; cell type; System; Performance; Informatics; validation studies; Sampling; Intervention Strategies; interventional strategy; Intervention; Genomics; cell preparation; Pharmaceutical Agent; Pharmaceuticals; Pharmacological Substance; Pharmacologic Substance; µfluidic; Microfluidics; Lysis; Cytolysis; Tissue Sample; Preparedness; Readiness; Address; Sequencing Core; DNA Sequencing Facility; Data; Regulatory Pathway; Reproducibility; Resolution; Resource Sharing; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Validation; Mammary Gland Parenchyma; Breast Tissue; Mammary Gland Tissue; Preparation; Characteristics; Resected; Process; neuron development; neuronal development; cost; novel strategies; new approaches; novel approaches; novel strategy; cost effective; Population; Consumption; transcriptomics; clinically relevant; clinical relevance; usability; preclinical evaluation; pre-clinical evaluation; flexibility; flexible; clinical investigation; web portal; internet portal; on-line portal; online portal; web-based portal; single cell technology; recruit; single-cell RNA sequencing; scRNA-seq; single cell RNA-seq; single cell RNAseq; single cell expression profiling; single cell transcriptomic profiling
