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CRISPR activation

Up-regulate gene expression, without cutting the DNA

CRISPR activation (CRISPRa) allows researchers to up-regulate specific gene function by activating transcription, without editing the DNA.

Our CRISPRa reagents provide a straightforward, efficient set of tools to study a gene's function.

Visit our CRISPRa applications page to learn more about key benefits to using CRISPRa for gene over-expression.

Reagents

CRISPRa guide RNA

CRISPRa dCas9-VPR

CRISPR SAM system

MS2 tracrRNA

For use with synthetic CRISPRa crRNA and a stable cell line expressing both components (dCas9-VP64 and MS2-p65-HSF1) of the SAM CRISPRa system.

Controls, additional reagents, and libraries

Turns genes on using programmable targeting - dCas9-based CRISPR systems can activate specific endogenous genes when guided to promoter/enhancer regions
Higher precision than conventional overexpression approaches - can modulate native gene regulation rather than relying on strong transgenes
Scalable for functional genomics - useful for pooled and arrayed CRISPR activation screens to systematically identify genes or pathways that drive phenotypes
More physiologically relevant gene expression - can better reflect endogenous regulatory context compared with artificial overexpression

CRISPRa reagent selection

There are many options and considerations for CRISPRa experimental conditions. Generally, users achieve the most robust activation when working with a stable population of dCas9-VPR expressing cells. However, many formats of transient and lentiviral delivery exist.

dCas9-VPR source	Guide RNA format	Delivery method	Recommendations for use & benefits
CRISPRa dCas9-VPR stable cells or CRISPRa dCas9-VPR lentiviral particle transduction & selection for dCas9-VPR stable cells	CRISPRa synthetic sgRNA	Transfection or electroporation	Short timepoint assays (2-4 days) Pool guide RNAs for increased activation Multiplex targets for simultaneous activation Arrayed screening
	CRISPRa synthetic crRNA + tracrRNA	Transfection or electroporation
	CRISPRa lentiviral sgRNA particles	Transduction	Extended timepoint assays with stable expression Low MOI - single integration for stable and even activation in cell population
CRISPRa dCas9-VPR mRNA	CRISPRa synthetic sgRNA	Co-transfection or electroporation	Short timepoint assays Antibiotic and fluorescent enrichment options No exogenous DNA, no integration of reagents in genome
CRISPRa dCas9-VPR mRNA	CRISPRa synthetic crRNA + tracrRNA	Co-transfection or electroporation
CRISPRa all-in-one dCas9-VPR + sgRNA		Transduction	Maximize successful delivery of CRISPRa components into difficult to transduce cell types Puromycin or GFP markers allow for live cell tracking of gene activation
Strict-R inducible lentiviral dCas9-VPR CRISPRa		Transduction	Reversible, small molecule-induced activation of target genes with tight regulation of gene expression Tet Degron stabilization for induction of dCas9-VPR expression upon the addition of doxycycline and Shield1, facilitating dynamic, time-controlled experiments
CRISPRa lentiviral and All-in-one lentiviral dual-sgRNA		Transduction	Deliver two sgRNAs targeting either the same gene or two different genes in a single lentiviral vector Lentiviral sgRNA and All-in-one lentiviral sgRNA formats available

As you explore CRISPR-mediated gene activation, consider whether the NEXTFLEX™ Rapid Directional RNA Kit 2.0 could complement your gene modulation analysis through additional transcriptome profiling.

CRISPRa workflows

CRISPRa workflow diagram with stable dCas9-VPR expression

CRISPR activation workflow with Lentiviral dCas9-VPR and synthetic crRNA:tracrRNA (right side) or synthetic sgRNA (left side).

CRISPR Modulation All In One

CRISPR activation workflow where either CRISPRa All-in-one lentiviral reagents or CRISPRa lentiviral sgRNA and dCas9-VPR lentiviral reagents can be transduced for activation in hard to transfect cell types.

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