Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

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

ACCESS DATA

Json Xml

Similar Datasets

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

Cas9 gRNA engineering for selectable genome editing, activation and repression

Project description:RNA-Seq after Cas9-gRNA transfection with different length gRNAs we performed PolyA Selection and RNA-Seq on cells transfected with dCas9-VPR and a gRNA of each length (20nt, 16nt, or 14nt) targeting ACTC1, MIAT, or HBG1/2

2015-07-15 | E-GEOD-70694 | biostudies-arrayexpress

Genome Editing with Tyrosinase gRNA

Project description:Genome Editing with Tyrosinase gRNA

| PRJNA937212 | ENA

Genome Editing with Tyrosinase gRNA

Project description:We confirmed Tyrosinase genome insertions or deletions of the three Tg mice by deep sequencing using MiSeq. These mice generated gene editing mice by using Tol2 transposon to introduce tyrosinase guide RNA (gRNA) into fertilized eggs obtained by crossing LSL (loxP-stop-loxP)-Cas9 mice with CAG-CreER mice.

2023-02-27 | GSE225711 | GEO

Cas9 gRNA engineering for selectable genome editing, activation and repression

Project description:RNA-Seq after Cas9-gRNA transfection with different length gRNAs

2015-07-15 | GSE70694 | GEO

RNASeq of Arg2 gRNA or scrambled gRNA CRISPR'd Tregs

Project description:RNASeq of Arg2 gRNA or scrambled gRNA CRISPR'd Tregs

| PRJNA588065 | ENA

Zebrafish 22 hpf embryos: WT vs. MO-grnA injected

Project description:Transcriptional profiling of zebrafish embryos comparing wild type untreated embryos with embryos injected with morpholino of zf-grna. This assay is used for the determination of expression profiling at 22 hpf under GRN-A deficiency. Two-condition experiment: wild type vs. MO-grnA treated cells. 3 biological replicates: each group contains 200 embryos.

2013-03-28 | E-GEOD-28318 | biostudies-arrayexpress

Optogenetic neural stem cells microarray

Project description:Optogenetic neural stem cells microarray

| PRJNA417074 | ENA

Generation of NPM1-ALK translocation in human T lymphocytes faithfully recapitulates the diversity of ALK+ ALCL tumor phenotype

Project description:To recapitulate the t(2;5)(p23;q35) chromosomal translocation, activated T lymphocytes from healthy donors PBMC were transfected with the RNP complex composed of the protein Cas9 with gRNA NPM1 (targeting NPM1 gene) and gRNA ALK (targeting ALK gene). Control cells were wild-type CD4+ activated lymphocytes. Cells were grown for 10-15 days in vitro prior to engraftment in vivo. Mice engrafted with NPM1-ALK+ T-cells developed T-cell lymphoma after 2-5 months. To get insights into the molecular bases of NPM1-ALK transformation, we next established the transcriptomes at an early in vitro stage and in fully transformed in vivo models. For this, we performed RNAseq analysis from 3 groups: 1-in vitro wild-type CD4+ activated lymphocytes [n=3], 2-in vitro NA-engineered CD4+ lymphocytes [n=3] and 3-in vivo derived CD4+ lymphoma cells purified by flow cytometry to distinguish CD3+ [n=3] and CD3- populations [n=3].

2022-02-09 | E-MTAB-10924 | biostudies-arrayexpress

Sensing Cardiac Electrical Activity with a Cardiomyocyte Targeted Optogenetic Voltage Indicator

Project description:Rationale: Monitoring and controlling cardiomyocyte activity with optogenetic tools offers exciting possibilities for fundamental and translational cardiovascular research. Genetically encoded voltage indicators may be particularly attractive for minimal invasive and repeated assessments of cardiac excitation from the cellular to the whole heart level. Objective: To test the hypothesis that cardiomyocyte-targeted voltage-sensitive fluorescence protein 2.3 (VSFP2.3) can be exploited as optogenetic tool for the monitoring of electrical activity in isolated cardiomyocytes and the whole heart as well as function and maturity in induced pluripotent stem cell (iPSC)-derived cardiomyocytes. Methods and Results: We first generated mice with cardiomyocyte-restricted expression of VSFP2.3 and demonstrated distinct sarcolemmal localization of VSFP2.3 without any signs for associated pathologies (assessed by echocardiography). Optically recorded VSFP2.3 signals correlated well with membrane voltage measured simultaneously by patch-clamping. The utility of VSFP2.3 for human action potential recordings was confirmed by simulation of immature and mature action potentials in murine VSFP2.3 cardiomyocytes. Optical cardiograms (OCGs) could be monitored in whole hearts ex vivo and minimally invasively in vivo via fiber optics at physiological heart rate (10 Hz) and under pacing-induced arrhythmia. Finally, we reprogrammed tail-tip fibroblasts from transgenic mice and used the VSFP2.3 sensor for benchmarking functional and structural maturation in iPSC-derived cardiomyocytes. Conclusions: We introduce a novel transgenic voltage-sensor model as a new method in cardiovascular research and provide proof-of-concept for its utility in optogenetic sensing of physiological and pathological excitation in mature and immature cardiomyocytes in vitro and in vivo. Determination of transgene (VSFP2.3) cardiotoxicity

2015-09-02 | E-GEOD-69190 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data