Methods for Generating Pooled RNA Oligos for Multiplexed Chromosome Targeting, Imaging, and Perturbation

Stanford researchers have developed a novel CRISPR-based method, Oligo-LiveFISH, for generating large-scale pools of synthetic RNA oligos that enable multiplexed targeting, imaging, and manipulation of genomic regions in living cells. This technology overcomes longstanding barriers in live-cell genomic imaging, particularly the challenge of visualizing non-repetitive DNA sequences, and opens new avenues for high-throughput functional genomics, drug discovery, and gene therapy research.

Traditional approaches for studying genome structure and function, such as FISH and 3C-based methods, are limited to fixed cells and cannot capture real-time chromatin dynamics. Existing CRISPR imaging tools are typically restricted to repetitive genomic regions and require laborious engineering or viral delivery, making them unsuitable for primary cells or high-throughput applications. Oligo-LiveFISH overcomes these barriers by using large pools of chemically synthesized RNA guides, which can be fluorescently labeled and delivered directly into cells as ribonucleoprotein complexes. This approach enables multiplexed tagging, imaging, and editing of many genomic regions at once, without the need for viral delivery or stable cell line creation. Oligo-LiveFISH has been demonstrated in a range of cell types, including human T cells and neurons, and can be adapted for use in diverse organisms.

Stage of Development: Research - in vitro