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Gene Knockout Kits

The most reliable CRISPR gene editing strategy, guaranteed. Intelligent gRNA design induces fragment deletions for knockout rates greater than 80%—shipping in as few as 5 days.

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Overview

Simple CRISPR knockout kits for any human or mouse protein-coding gene

  • Faster Data: Discover faster and eliminate the trial and error of guide screening. Get your Gene Knockout kit delivered in 5 days.
  • Efficiency: The most reliable knockout strategy. High knockout efficiency, guaranteed.
  • Simplified Online Ordering: Easily order online through our ordering portal. Complete your experiment with controls, SpCas9 nuclease and Transfection Optimization kit 
Gene Knockout Kits

Human and Mouse Knockouts Have Never Been Easier.

Common CRISPR knockout strategies rely on an individual guide RNA (gRNA) that produces random indels in the gene of interest. This approach produces a single double-strand break at the targeted locus and relies on the cellular repair mechanism to generate a frameshift mutation to disrupt the gene. This strategy is inefficient due to the wide variety and unpredictability of edits that can occur and often does not generate a functional knockout.

EditCo’s Gene Knockout Kit was designed to increase the efficacy of CRISPR-mediated knockouts. Typical guide RNA strategies rely on the generation of indels, which does not always result in complete gene disruption. EditCo’s multi-guide approach consistently generates fragment deletions at the targeted loci, so you can be confident in your gene knockout. 

Multi-Guide Schematic

 

Deliverables

Gene Knockout Kits for any human or mouse genes

Our Gene Knockout Kit is guaranteed for human and mouse (excluding non-essential genes).
The multi-guide sgRNA is chemically modified to resist degradation and prevent triggering intracellular immune responses.

Complete your CRISPR experiment with the Transfection Optimization Kit, add-on controls and SpCas9 nuclease.

Features

Species
  • Human
  • Mouse
Sizes
  • 1.5, 3, 5, 10 nmol
Format
  • Individual tubes

Deliverables

  • nmol target-specific multiguide sgRNA, tube, dry 
  • sgRNAs are modified: 2' O-Methyl analog on first and last 3 bases; 3' phosphorothioate between first 3 and last 2 bases (modifications help resist degradation and prevent triggering intracellular immune responses).
  • SpCas9 proprietary EZ Scaffold
  • Nuclease-free water 1.5 ml (tube) (1 for every 5 kits)
  • TE Buffer 1.5 ml (tube) (1 for every 5 kits)
  • QC document (pdf) and sgRNA and Primer sequences for PCR and Sanger sequencing .csv file
Data

Smart Guide Design for Guaranteed Knockouts

Multi-guide Design Consistently Induces Deletions

Multiguide gRNA approach

Figure 1. EditCo’s Gene Knockout Kit was designed to increase the efficacy of CRISPR-mediated knockouts. The multi-guide approach consists of 3 gRNAs spatially coordinated to create a fragement deletion in a single exon. Targeting a single early exon of a gene induces multiple concurrent double-strand breaks in the target sequence of a gene. This results in disruptive one or more >21 bp fragment deletions. This approach is associated with higher frequencies of knockout alleles in edited pools but reduced occurrence of off-target edits as compared to single-guide approaches.

 

Multiguide gRNA
Figure 2. Multi-guide gRNA for each target resulted in >75% large deletion outcomes. Individual vs. multi-guide gRNA editing was compared for 2 gene targets (TNF, TLR4) in dendritic cells (transfected via nucleofection). Editing efficiency was analyzed by sequencing the targeted loci on a MiSeq and sequencing outcomes were categorized based on editing type (no indel, large deletion ≥50bp, small deletion <50bp, insertion). 

 

Consistent and Reliable Knockouts

 

EB Libraries Multiguide Scatter

Figure 3. Better knockout efficiency was found across 32 genetic targets assessed. Guide RNAs displayed 29.2% better median knockout efficiency when introduced in a multiguide format (89.9% KO score) relative to an individual sgRNA format (69.6% KO Score).

 

Fragment Deletions Result in Sustained Protein Depletion

Fragment deletions caused by multi-guide sgRNA editing result in protein knockout after seven days post-transfection in non-clonal cell lines.

Editing of three genes in HEK293 cells

Figure 4. Editing of three genes (CDK9, CINP, COASY) in HEK293 cells (transfected via nucleofection) using the multi-guide approach resulted in high knockout efficiencies, as indicated by Knockout (KO) Scores. Western blots showed loss of the corresponding proteins relative to mock treated cells (no sgRNA or Cas9). For seven days post-transfection, cells were assessed for the presence of the protein target via immunoblot analysis (GAPDH expression across the same timeframe was used for normalization).

 

multi-guide KO gel-1

Figure 5. Western blot analyses for all 5 knockout target genes indicate a complete depletion of proteins across the 3 specified time points (days 7, 14, and 21) relative to the negative controls. Knockout cell pools for the target genes were generated by nucleofecting U2OS cells with multi-guide sgRNA and Cas9 (as RNPs). A negative control pool was also transfected for each target using non-targeting sgRNA.

Resources

Application Note
Multi-guide RNA Improves CRISPR Efficiency by Generating Fragment Deletions
Learn how intelligently designed multiple gRNAs improve CRISPR knockout efficiency with single exon fragment deletions.
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E-book
Cell Engineering 101 eBook
Discover methods and insights into skillfully generating knockout or knock-in CRISPR edits in a wide-range of cell lines.
Learn More
flyer
Gene Knockout Kit: Elevate Your Guide Design Strategy
EditCo's Gene Knockout Kit takes the trial and error out of CRISPR knockouts by using a novel strategy of guide RNA design to knock out any human or mouse protein-coding gene.
Learn More
Quick Start Guide
Gene Knockout Kit Quick Start Guide
This quick-start guide can be used for EditCo's Gene Knockout Kit. In this guide, we include important information on how to store, quantify, and transfect your gRNA kit and analyze your knockout kit experiments.
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EditCo's Automated CRISPR Platform

Integrating our core CRISPR expertise, high-quality reagents, and automated processes, we deliver the best edited cell-based models at any scale.

EditCo Reagent Workcell
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Leveraging a sophisticated infrastructure with integration between bioinformatics, software, and automated platforms
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Automating biology and synergistic disciplines for streamlined cell editing and cell culture workflows
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Delivering experiment-ready edited cells for your next discoveries through a robust and cohesive ecosystem
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Principal Investigator, Boston Children's Hospital
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Albert Rose, Ph. D.
Principal Investigator, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School
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Albert Rose, Ph. D.
Principal Investigator, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School
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Albert Rose, Ph. D.
Principal Investigator, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School
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