In order to meet the investigative needs of the Dartmouth research community in genomics, a request of $175,000 is submitted as part of a proposal to purchase a Chromium instrument from 10X Genomics, Inc. (Pleasanton, CA) to carry out single-cell sequencing. The sequencer will be placed in the Dartmouth Genomics & Molecular Biology Shared Resource (GMBSR), a core facility (1500 sq ft) in the Norris Cotton Cancer Center (NCCC) at the Geisel Medical School at Dartmouth with the expertise, knowledge, and experience to maximize the scientific potential of the new technology. The Chromium system would provide users with the latest and most advanced high-throughput sequencing technology available for single cell genomics. The need for single cell sequencing is based on several factors. Foremost, the provision of this new technology will allow Dartmouth investigators to probe questions in cancer biology, immunology, and other disciplines in ways not possible with the current technology at Dartmouth, including questions regarding the high degree of cell-to-cell variability in cancer progression and resistance to therapy, the immune response, and chronobiology. A user group of Dartmouth PIs has been identified of which seven major users and several minor users are projected to use the new technology immediately, with additional users predicted in the near and sustainable future. The major and minor user groups possess 11 NIH grants totaling approximately $4.34 million in annual direct costs. The single cell sequencing would be secured in an existing pipeline consisting of the (i) Immune Monitoring and Flow Cytometry Shared Resource to advise, assist, and/or carry out the labeling, isolation, and verification of single cell preparations; (ii) the above described GMBSR to carry out the single cell library construction and deep sequencing; and (iii) Bioinformatics resources to advise, assist, and/or carry out the analyses. The personnel, expertise, and resources to successfully operate and sustain single cell sequencing are already in place. The pipeline currently operates under the auspices and support of the NCCC with additional support from the Geisel Medical School at Dartmouth and Dartmouth College. Administration and scientific issues regarding the technology will be overseen by the Integrative Biology (Advisory) Committee. Operating costs will for the most part be met by subsidies from an NCI Cancer Center grant to the NCCC and fee-for-service charges to the investigator. In conclusion, the investigators, research questions, infrastructure, and cross-institutional and -departmental support are in place to implement single cell sequencing to greatly advance genomics studies and knowledge at Dartmouth.
The 10X Genomics Chromium system is at the cutting edge of high-throughput single cell genomics technology. The technology will be used as an integral part in the research efforts of dozens of investigators at Dartmouth. The use of the single cell sequencing technology will therefore promote genomics research at Dartmouth in understanding basic biological questions and the basis of different diseases and in the search for more efficacious therapeutic approaches for those diseases.