Tag: VP64

Specific Modulation of CRISPR Transcriptional Activators through RNA-Sensing Guide RNAs in Mammalian Cells and Zebrafish Embryos

Modular iSBH-sgRNA designs enable spatial separation of spacer and trigger-sensing sequences. A. In second-generation iSBH-sgRNAs, RNA triggers are complementary with the iSBH-sgRNA backfolds, thus sgRNA spacers influence RNA trigger sequences. In modular iSBH-sgRNAs, design constrains were eliminated as triggers are only complementary with the iSBH-sgRNA loop and first 15nt of…

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Implanted cells triggered by electricity used to drive in vivo gene expression

Design of the direct-current-activated transgene expression switch in mammalian cells. a, Schematic illustration of the stimulation setup for monolayer cultures. Each well of a 24-well plate has two platinum wires that function as anode and cathode, placed 0.6 cm apart submerged in the culture medium. When electric current is applied, bubbles…

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IJMS | Free Full-Text | Engineering Human Cells Expressing CRISPR/Cas9-Synergistic Activation Mediators for Recombinant Protein Production

1. Introduction Peptides are employed for a multitude of therapeutic approaches and can be obtained from the native organ or tissue in which they are produced as well as through recombinant methods where nucleic acids encoding a candidate gene(s) are delivered to and expressed in cells in vitro. As an…

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Targeted Transcriptional Activation in Human Cells and in the Mouse Brain Tissue by Capsid-modified AAV and Evolved CRISPRa – CityU Scholars

Transcriptional dysregulation has been implicated in a number of progressive neurodegenerative diseases. To correct transcriptional misregulation, emerging CRISPR tools such as CRISPR activation (CRISPRa) and CRISPR interference (CRISPRi) could be used. In addition, efficient and precise transcriptional activation in broad brain areas and multiple neuroanatomical structures can be achieved by…

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CRISPRa experiment with a silent sgRNA at the LPL locus does not influence cell proliferation nor the expression of senescence marker genes.

(A) teloHAEC that express dCas9-VP64 were infected with a lentivirus that carries a sgRNA that targets rs1441755 at the LPL locus. This sgRNA was silent in all our pooled CRISPR screens for all six endothelial phenotypes tested. In the absence or presence of antibiotic selection (Zeocin), LPL_sg08237 does not affect…

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ZFIN Publication: Weuring et al., 2021

PUBLICATION Authors Weuring, W.J., Dilevska, I., Hoekman, J., van de Vondervoort, J., Koetsier, M., van ‘t Slot, R.H., Braun, K.P.J., Koeleman, B.P.C. ID ZDB-PUB-210819-5 Date 2021 Source The CRISPR journal   4: 575-582 (Journal) Generate reference Registered Authors Koeleman, B. P. C., van ‘t Slot, Ruben, Weuring, Wout Keywords none…

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Nanoscale, antigen encounter-dependent, IL-12 delivery by CAR T cells plus PD-L1 blockade for cancer treatment | Journal of Translational Medicine

Lentiviral vector construction and production The lentiviral vector was designed as previously described [13, 41]. LdCV vector contains the following: human codon-optimized S. pyogenes dCas9 which was fused at the C-terminus with VP64-p65-Rta (VPR). LAT-TCS-dCas9-VPR was assembled by fusing LAT (Human cDNA, NM_001014987.2) with dCas9-VPR-Q8 and cloned into a modified…

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dCas9-VP64-Blasticidin SAM CRISPRa Helper Construct 1 Plasmid DNA

This product is a lentiviral plasmid that utilizes the EF1 alpha promoter to drive expression of dCas9-VP64 and blasticidin resistance cassette linked by a 2A peptide (EF1a-dCas9-VP64-2A-Blasticidin) allowing for easy selection following successful transfection or transduction. Use Sigma′s lentiviral dCas9-VP64 plasmid for generation of lentiviral particles and efficient production of…

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