CRISPR-SNIPER Gene Editing Services

Overview

CRISPR-based gene editing technology is revolutionizing the biopharma industry, enabling advances in disease modeling that were impossible just a few years ago. CRISPR-Cas9 enables precise, directed creation of knock-outs and knock-ins (including single base changes) in all types of cells. REPROCELL provides CRISPR-Cas9-based gene editing in collaboration with GenAhead Bio.

Our technology uses the proprietary SNIPER method to screen for positive clones. This streamlines the most time-consuming part of the CRISPR-based gene editing: Screening colonies for positives.

Please visit CRISPR-SNIPER Example Projects to see CRISPR-SNIPER gene editing in action.

 

CRISPR-SNIPER Gene Editing

  • Based on the gold-standard CRISPR-Cas9 gene editing system
  • Includes SNIPER clone screening technology for accelerated identification of positive clones
  • Delivers high success rate for single base pair change modifications for disease model generation
  • Great for knock-out (KO) and knock-in (KI) applications
  • Provides Freedom-to-Operate for research use including drug screening

CRISPR-Cas9 Gene Editing Complex

CRISPR-Cas9 Cartoon


 

Traditional knock-in generation uses a screening gene, such as PurR or GFP to aid in identifying positive. For single base change generation, use of screening genes is not possible, since only the desired base change is. This means that screening of large numbers of colonies is required to identify positive clones, resulting in a significant investment in time and resources. Addition of SNIPER to single base change modification screening results in up to a 100-fold or greater increase in fraction of positive clones, greatly speeding this resource-intensive step.

Comparison of Screening Methods

ConditionPositive Clones
SNIPER Screening20-30+%
Traditional Screening0.1-1%

 

What is SNIPER?

SNIPER (Specification of Newly Integrated Position and Exclusion of Random-integration) uses a combination of long, high-stringency donors with a dPCR-based prescreen to enable focus on colonies most likely to be positive. This allows for increased success rate in isolating homoallelic and heteroallelic clones with the correct genetic change, which would be difficult to isolate using conventional methods. For more information, please see our recent webinar.

 

CRIPR-SNIPER Gene Editing Project Outline

 Milestone 1 ▼Milestone 2 ▼Milestone 3 ▼
 Bulk Screening for optimal conditionScheduled CloningCell Expansion and QC
Scope of Work
  • Optimization of targeting guides
  • Set optimal conditions
  • Transfection
  • Bulk screening
  • Selection
  • Colony Pick-up
  • 2nd Screening
  • Expansion
  • QC
  • Cryo preservation in vials
  • Delivery

Check Points
  • Confirm Transfection condition by bulk screening with PCR
  • Verify success in clonal cells with PCR
  • Sequence Verification
  • Expression of undifferentiated marker
  • Mycoplasma

CRISPR-SNIPER projects, like all of our service projects, are developed in close collaboration with our clients and are organized around milestones to minimize risks.

 

Our experience

Types of projects

Types of projects

 

 

Our gene editing team is experienced in creating a variety of types of mutants including challenging Marker Insertions and SNP (single base change) creations.

We have worked with a variety of cell types including iPS Cells and adherent and suspension cancer cells.


Please visit CRISPR-SNIPER Example Projects to see CRISPR-SNIPER gene editing in action.

For more technical information, please see our webinar: CRISPR/Cas9 using SNIPER Technology — Managing Challenging Cases

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