This technology is a method for optical in situ nucleotide sequence detection using combinatorial hybridization.
Genetic screens enable the study of various genes underlying specific cellular phenotypes. However, screening is often limited due to issues with scalability, cost, and acquiring correct and adequate resolution and readout, as well as the lack of methods that simultaneously read out transcriptomic, proteomic and perturbation identity in single cells without destruction. One approach to circumvent these limitations is the use of optical pooled screening techniques; however, even these approaches are limited by difficulties in imaging complex cellular phenotypes.
This technology is a method for CRISPR screening that employs cyclic hybridization-based barcode readout, allowing for in situ nucleotide sequence detection within cells. This method utilizes paired primer and padlock detection oligos that require hybridization to their respective target sequences, allowing the primer to serve as a template for the circularization of the padlock probe. Together, these features enable robust spatial and temporal analysis of cell cultures, primary and stem cells, and tissue sections. This pooled approach can be used to study hundreds to thousands of targets simultaneously, enabling large-scale studies while reducing costs and permitting higher throughput screening and resolution. This technology has been validated using human breast cancer cell lines, fibroblasts, hESCs, iPSCs, motor neurons, and tumor xenograft tissue.
Patent Pending(US19/401,747)
IR CU24118
Licensing Contact: Joan Martinez
