Under the leadership of Dr. Keji Zhao and the steering committee (Drs. Mark Knepper, Warren Leonard, Chris O'Donnell, Adrian Wiestner), the DIR DNA Sequencing and Genomics Core (DSGC) has been providing a wide spectrum of high-throughput genomics services to facilitate basic and translational research at NHLBI and across the NIH. (1) Consultation and data acquisition: the DSGC has provided the state-of-the-art genomics services for DIR investigators in a cost effective and timely fashion. Diverse projects have been carried out including whole-genome sequencing, exome sequencing, transcriptome sequencing (RNA-seq and small RNA sequencing), ChIP-seq, PAR-CLIP, RIP-Seq, ChIA-PET, Mito-seq, RCA-seq, as well as several innovative NGS applications (e.g. TSS-seq, PA-seq) to interrogate genome/transcriptome diversity. (2) In-depth data analysis: The DNA Sequencing and Genomics Core has explored and implemented a wide range of open source and commercially available software packages for primary and secondary NGS data analysis. In addition, project-specific data analysis is further achieved by in-house software and algorithm development. As the result, it provides a broad spectrum of tools for DIR investigators to convert the high-throughput data into biological meaningful findings for further examination. (3) Training and education: The DSGC has taken diverse venues to promote broad dissemination of NGS technology. Consultations are provided for experimental design and data analysis. The core also offers routine one-on-one training for library preparation and data analysis. (4) Research and development: Several R&D projects were carried out in collaboration with the DIR investigators. High priority was set for technologies that are beneficial to multiple users or expected to facilitate the broad applications of high-throughput sequencing platform. We have developed and optimized protocol for (a) mitoRCA-seq to identify single nucleotide mutations and indels related to aging and heart diseases (b) targeted sequencing assays for myeloid leukemia, pheochromocytomas, congenitial heart disease (c) single-cell transcriptome analysis with Fluidigm C1 platform and an improved SPA-seq procedure. In addition, we have been developing computational pipelines and workflow for systematic identification of SNPs, alternative polyadenylation, global intron retention and other regulatory events in gene expression network.

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Budget End
Support Year
6
Fiscal Year
2015
Total Cost
Indirect Cost
Name
U.S. National Heart Lung and Blood Inst
Department
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