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Light induced expression of gRNA allows for optogenetic gene editing of T lymphocytes in vivo

ABSTRACT: Light induced expression of gRNA allows for optogenetic gene editing of T lymphocytes in vivo

PROVIDER: PRJNA1152922 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30413000_1.fastq.gz	Fastqsanger.gz
	SRR30413000_2.fastq.gz	Fastqsanger.gz
	SRR30413001_1.fastq.gz	Fastqsanger.gz
	SRR30413001_2.fastq.gz	Fastqsanger.gz
	SRR30413002_1.fastq.gz	Fastqsanger.gz

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Light induced expression of gRNA allows for optogenetic gene editing of T lymphocytes in vivo

Project description:There is currently a lack of tools capable of perturbing genes in both a precise and spatiotemporal fashion. The flexibility of CRISPR, coupled with light's unparalleled spatiotemporal resolution deliverable from a controllable source, makes optogenetic CRISPR a well-suited solution for precise spatiotemporal gene perturbations. Here we present a new optogenetic CRISPR tool (BLU-VIPR), that diverges from prevailing split-Cas design strategies and instead focuses on optogenetic regulation of gRNA production. We engineered BLU-VIPR around a new potent blue-light activated transcription factor and ribozyme-flanked gRNA. The BLU-VIPR design is genetically encoded and ensures precise excision of multiple gRNAs from a single mRNA transcript This simplified spatiotemporal gene perturbation and allowed for several types of optogenetic CRISPR, including indels, CRISPRa and base editing. BLU-VIPR also worked in vivo with cells previously intractable to optogenetic gene editing, achieving optogenetic gene editing in T lymphocytes in vivo.

2025-03-21 | GSE275757 | GEO

Light-induced expression of gRNA allows for optogenetic gene editing of T lymphocytes in vivo.

Project description:There is currently a lack of tools capable of perturbing genes in both a precise and a spatiotemporal fashion. The flexibility of CRISPR (clustered regularly interspaced short palindromic repeats), coupled with light's unparalleled spatiotemporal resolution deliverable from a controllable source, makes optogenetic CRISPR a well-suited solution for precise spatiotemporal gene perturbations. Here, we present a new optogenetic CRISPR tool (Blue Light-inducible Universal VPR-Improved Production of RGRs, BLU-VIPR) that diverges from prevailing split-Cas design strategies and instead focuses on optogenetic regulation of guide RNA (gRNA) production. We engineered BLU-VIPR around a new potent blue-light activated transcription factor (VPR-EL222) and ribozyme-flanked gRNA. The BLU-VIPR design is genetically encoded and ensures precise excision of multiple gRNAs from a single messenger RNA transcript. This simplified spatiotemporal gene perturbation and allowed for several types of optogenetic CRISPR, including indels, CRISPRa, and base editing. BLU-VIPR also worked in vivo with cells previously intractable to optogenetic gene editing, achieving optogenetic gene editing in T lymphocytes in vivo.

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