HiTS Seminar: David Feldman, 10:00 am, Thursday, January 17, 2019
Broad Institute, Cambridge, MA
Scalable Image-based Genetic Screens
Genetic screens (e.g., CRISPR loss-of-function, ORF gain-of-function) provide a powerful means to identify genes underlying cellular processes. A wide range of cell behaviors can be observed by imaging assays, including dynamic responses from the scale of minutes to generations. However, image-based screens have been difficult to scale due to the requirement to test each perturbation one at a time. We developed a pooled approach for image-based genetic screens in mammalian cells, using in situ sequencing to sequence the perturbation (e.g., CRISPR sgRNA) directly in fixed cells. We piloted the method by imaging live-cell NF-kB translocation kinetics in a collection of ~1,000 gene knockouts and are now moving to genome-scale. We hope that this approach, which requires no special hardware beyond an automated widefield microscope, will enable widespread use of highly multiplexed image-based screens.
10:00 am – 11:00 am
Thursday, January 17, 2019
Harvard Medical School
200 Longwood Avenue
Warren Alpert Bldg, 5th Floor, Room 563
Boston, MA 02115
Hosted by Michael Tsabar, Ph.D.