Year: 2025
Type: article
Abstract: The biggest challenge in using CRISPR technologies, which limits their widespread application in medicine, is off-target effects. These effects could, in principle, be minimized by ensuring that CRISPR is activated primarily in the targeted cells, thereby reducing the likelihood of unintended genetic modifications in non-target tissues. Therefore, the development of a light activatable CRISPR approach to dynamically control gene activation in both space and time would be highly beneficial. A drawback is that the overwhelming majority of recently introduced light activatable CRISPR systems require UV or blue light exposure, severely limiting the penetration depth of light in tissue at which CRISPR can be activated, and, in the case of UV light, raising safety concerns. A small number of systems that activate CRISPR using longer wavelengths are hindered by either slow light activation or issues related to toxicity and biocompatibility of the proposed techniques in humans. To address this, we developed a split-Cas9/dCas9 system in which activation is achieved through a near-infrared photocleavable dimerization complex. This photoactivation method can be safely used in humans in vivo, easily adapted to different split-Cas9/dCas9 systems, and enables rapid, spatially precise light activation across various cell types.
Source: Light Science & Applications
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $4,190
Funders: National Science Foundation, Foundation for the National Institutes of Health, National Institutes of Health, National Institute on Aging, National Cancer Institute, National Institute of Biomedical Imaging and Bioengineering, Division of Chemical, Bioengineering, Environmental, and Transport Systems, Division of Emerging Frontiers in Research and Innovation
Awards: G3369319967, G3614377485, EFRI CEE: Optically Controlled Localized Epigenetic Chromatin Remodeling with Photoactivatable CRISPR-dCas9, G5091354950, G5954878796, G5994914234, Biosensing of chromatin packing in live cells, Broadband multispectral microscopy of insulin granule dynamics in live pancreatic islets, G6862189080, G7052197093, G7131850347, Multispectral imaging of bile duct epithelial morphology for early detection of malignancy, A lab-on-a-chip device for measuring multiple biochemical and biophysical biomarkers of Alzheimer’s disease, G8051096807, Glaucoma Epigenetic Treatment with Light Activated CRISPR-dCas9, G9000147263, G9000281343
OpenAlex ID: W4411911608