Essentialome-Wide Multigenerational Imaging Reveals Mechanistic Origins of Cell Growth Laws.

ABSTRACT: Optical Pooled Screening (OPS) enables precise measurement of pooled genetic libraries in single, highly multiplexed microscopy experiments, ^1-4 linking image-based phenotypes to genotypes at genome-scale. However, no method has combined large-scale OPS with multi-generational time-lapse imaging, which is especially important for studying single-cell dynamics and physiology in microbes. Here we develop MARLIN, a technique for multigenerational time-lapse OPS in a microfluidic device and leverage it to map live-cell imaging data to genetic perturbations in a ∼130,000-strain mismatch-CRISPRi library targeting all essential genes in E. coli . The resulting dataset comprises nearly a hundred million cell cycles and 1.6 billion cells, representing an average of 33 different kn

SUBMITTER: Eaton DS 

PROVIDER: S-EPMC12262249 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature