2022年2月7日 · Here, we describe a protocol for the design, execution and evaluation of such gene therapies using dCas9-VPR. We offer guidelines on how to identify functionally equivalent genes, design and...

Plasmid SP-dCas9-VPR from Dr. George Church's lab contains the insert SP-dCas9-VPR and is published in Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312. This plasmid is available through Addgene. More than 20 requests

2015年3月2日 · We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9.

2020年9月18日 · Here, we report a novel multifunctional CRISPR system that enables simultaneous gene activation, repression, and editing (CRISPR-ARE) with a single Cas9-VPR protein for combinatorial metabolic engineering applications in Saccharomyces cerevisiae. Via gRNA engineering, we achieved orthogonal transcriptional regulations and genome editing using ...

The fusion providing the highest transcriptional activation was VP64-p65-Rta, VPR for short. dCas9-VPR performs significantly better than dCas9-VP64, any of the single ADs fused to dCas9, and constructs containing double AD fusions. 13 dCas9-VPR can activate transcription of endogenous genes in animal cell lines, in some cases to levels ...

2023年3月11日 · Current CRISPR-activation (CRISPRa) platforms often require multiple components because of inefficient transcriptional activation. Here, we fused different phase-separation proteins to dCas9-VPR (dCas9-VP64-P65-RTA) and observed robust increases in transcriptional activation efficiency.

2020年8月19日 · Using dual adeno-associated viral vectors expressing split dCas9-VPR, we show efficient transcriptional activation and long-term expression of cone photoreceptor-specific M-opsin (Opn1mw) in a rhodopsin-deficient mouse model for retinitis pigmentosa.

We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. We demonstrate its utility in activating endogenous coding and noncoding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced ...

Here, we describe a protocol for the design, execution and evaluation of such gene therapies using dCas9-VPR. We offer guidelines on how to identify functionally equivalent genes, design and clone single guide RNAs and evaluate transcriptional activation in vitro.

Deactivated Cas9-VPR solutions for the CRISPR activation system The Horizon CRISPRmod CRISPR activation (CRISPRa) system is an adapted CRISPR-Cas9 system that is used for upregulation of genes. It utilizes a nuclease-deactivated S. pyogenes Cas9 (dCas9), that is fused to transcriptional activation domains (VP64, p65 and Rta).