Science Advances · 2021 · 44 citations · 57 references
Life SciencesBiotechnologyGeneticsGenetic EngineeringGenome EditingGenome EngineeringCrispr-based EngineeringCrispr-cas SystemsTarget Site SelectionCrispr-based Genome EngineeringCrispr-cas Genome EditingCrispr-cas RegulationCrispr-cas12 SystemsIn Vivo Genome Editing
The CRISPR-Cas12a has been harnessed as a powerful tool for manipulating targeted gene expression. The possibility to manipulate the activity of CRISPR-Cas12a with a more precise spatiotemporal resolution and deep tissue permeability will enable targeted genome engineering and deepen our understanding of the gene functions underlying complex cellular behaviors. However, currently available inducible CRISPR-Cas12a systems are limited by diffusion, cytotoxicity, and poor tissue permeability. Here, we developed a far-red light (FRL)–inducible CRISPR-Cas12a (FICA) system that can robustly induce gene editing in mammalian cells, and an FRL-inducible CRISPR-dCas12a (FIdCA) system based on the protein-tagging system SUperNova (SunTag) that can be used for gene activation under light-emitting diode–based FRL. Moreover, we show that the FIdCA system can be deployed to activate target genes in mouse livers. These results demonstrate that these systems developed here provide robust and efficient platforms for programmable genome manipulation in a noninvasive and spatiotemporal fashion.
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A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
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