Connor Tou (COLLINS & KLEINSTIVER LABS)

Precision Genome Editing with DNA-dependent Polymerases and HUH Endonucleases

Genome editing technologies continue to enable new therapeutic approaches. Here, we develop click editors (CEs), which synergize the advantageous properties of DNA-dependent polymerases, HUH endonucleases (HUHes), and RNA-guided nickases to install a range of edit types and lengths into the human genome without requiring double-strand breaks. CEs leverage the “click”-like ssDNA-protein attachment biochemistry of HUHes to covalently localize user-specified editing templates to targeted genomic loci for DNA-dependent polymerization. Through iterative optimization of the modular CE components, we demonstrate efficient and specific human genome editing, promising broad utility across diverse biological applications.

John Day (BOYER LAB)

HiExM: High-throughput Expansion Microscopy Enables Scalable Super-resolution Imaging

Expansion microscopy (ExM) enables nanoscale imaging using a standard confocal microscope through the physical, isotropic expansion of fixed immunolabeled specimens. Although ExM has been widely adopted, current methods cannot be performed in multi-well cell culture plates which limits the number of samples that can be processed simultaneously. We developed High-throughput Expansion Microscopy (HiExM), a robust platform that enables expansion microscopy of cells cultured in a standard 96-well plate. We employ HiExM to study endosomal organelles, using the augmented resolution afforded by expansion to understand organelle functional identity through organelle morphology.