The Dharmacon™ All-in-one CRISPR platform

Single reagent, easy delivery, straightforward selection

 

CRISPR workflows are simplified with Dharmacon All-in-one reagents. The All-in-one platform combines the CRISPR nuclease and single guide RNA expression into a single lentiviral packaged vector – for the simplest gene knockout/modulation workflow available.
One reagent CRISPR for any human or mouse gene.

All-in-one benefits:

  • Single reagent – CRISPR nuclease + sgRNA from one vector
  • Easy, ready-to-use – Simply thaw and add lentivirus to your cells, no transfection or electroporation necessary
  • Simple selection – Choose antibiotic resistance or fluorescent selection markers 
  • Our All-in-one system gives efficient gene knockout, activation, or interference

Ideal for:

  • New CRISPR users – simplified workflow for fast results
  • Easily generating cell models to study gene function
  • Difficult-to-transfect cells – Lentiviral delivery can be used with nearly any cell type
  • Hit validation – follow-up target hits from pooled-lentiviral screens

 

The DharmaconTM All-in-one system

The All-in-one system offers the simplest CRISPR workflow available – a single reagent, easy delivery and straightforward selection.

 
Single reagent

Using a single vector to express both CRISPR nuclease and gene-specific guide RNA eliminates the need to perform sequential transduction and selection steps.

 
Easy delivery

Lentiviral packaging allows for easy transduction into nearly any cell type. Eliminating the need for toxic transfection reagents or electroporation techniques makes the All-in-one system particularly useful for genome editing and transcriptional modulation in difficult-to-transfect or primary cell types.

Transduction-ready reagents also eliminate any cloning or in vitro transcription steps.

 
Straightforward selection

The choice of a puromycin resistance gene or EGFP marker allows for straightforward selection of cells that have successfully integrated the vector. EGFP is recommended for rapid enrichment of targeted cells, as FACS analysis may be performed as soon as fluorescence is expressed.

 

Promoter options

Choice of promoters allows you to select the most active promoter in your cell line.

 all-in-one vector maps 

 

All-in-one CRISPR gene knockout, activation, and interference workflow

 

 

The power of the All-in-one system lies in the simplicity of it's workflow

To transduce, simply thaw the reagent and add the appropriate amount to your cells. Then easily select for targeted cells using puromycin or enrich using FACS. 

When using puromycin selection, growth medium is replaced with puromycin selection medium 24-48 hours post-transduction. Cells are then monitored daily until growing normally in the selection medium. Once growth is stable, isolation and/or expansion of the targeted cell population(s) may occur.

Alternatively, using the EGFP fluorescent reporter option offers the ability to enrich for edited cells via FACS selection in as few as 72 hours post-transduction. The FACS enrichment workflow is especially useful for short-lived cell types, such as primary cells.

For CRISPRa and CRISPRi, we recommend the mixed cell population workflow.