L2M NSERC - Integrated technology for automated cell-by-cell genetic screening

点击翻译按钮获取中文摘要

The genomic information inside a cell (the genome) maps the structural and functional capabilities of a cell. With the advent of more sophisticated gene editing tools (e.g., CRISPR), it is possible to alter the genome by adding, removing or modifying DNA at a particular location with the hopes of obtaining cells with a desired function. By selecting a single cell with a unique genome through rounds of screening, it is possible to select for strains that are capable of producing valuable products (e.g., cannabinoids or biofuels). For more advanced cell characterization, it is necessary to isolate and to grow cells individually, as monoclones. Currently there are very few technologies available to perform the selection and the isolation. The main problems with current methods for generating and isolating the clones are (i) the high costs, (ii) lengthy steps, (iii) lack of control over cell sorting parameters and (iv) the absence of integrated methods for transferring the clones to well-plates, risking the loss of selected clones. In the Shih lab, we have created an invention called "D2-isolate", which is a hand-held device that integrates two microfluidic paradigms: droplet-in-channel and digital microfluidics (DMF). Droplet-in-channel microfluidics can operate in high-throughput ranges to generate droplets containing 0 or 1 cell, but only results in a large mixture of droplets to be analyzed while only ~10% of them contain a cell. DMF systems can address each droplet individually. The integration of the two enables the generation of droplets containing 1 cell with high efficiency (no empty droplet) while controlling/moving each droplet containing a single cell independently. Based on pre-programmed criteria, a cell can automatically be either discarded, or transferred to a 96-well plate for further characterization. As a fully integrated and automated platform, D2-isolate will expedite the workflow for applications related to single-cell generation and analysis. Hence, in this proposal, we aim to evaluate the market for our technology with the goal of expanding the application space related to single-cell technologies and help prepare our efforts towards commercialization.