CRISPR Reagents

Streamline your CRISPR with our intuitive, flexible ordering portal.

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

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.

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

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.

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.

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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.

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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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High Knockout Efficiency: Multi-guide design that results in functional knockouts of your targets—reliable results, guaranteed
Robust Quality Control: Consistently high quality and precision when plating provide you with reliable knockout results across your entire screen.
Unrivaled Delivery Speed: Faster turnaround time and consistent delivery means you get to your experiments quicker.

Screen Smarter Not Harder. Effective Knockout Strategy for Better CRISPR Screening.

Other loss-of-function screening libraries are often plagued with laborious preparation, data analysis, poor editing efficiency and low quality reagents that can be cytotoxic to more sensitive cell types such as primary cells. Our Arrayed CRISPR gRNA Libraries are different.

Powered by EditCo’s unique multi-guide design, our Arrayed CRISPR gRNA Libraries are comprised of SpCas9 sgRNAs targeting one gene per well in a multiwell plate format. The multi-guide design consists of up to three spatially coordinated SpCas9 sgRNAs that induce fragment deletions in an early single exon of a gene. Large fragment deletions in the target gene increased likelihood of functional knockout by not relying on random indel creation. Overall, this decreases the likelihood of false negatives, reliably knocking out any human or mouse protein-coding gene.

Our libraries are ready-to-transfect, either complexed into RNPs or directly into a Cas9-expressing cell line avoiding long preparation protocols. The arrayed format avoids the need of messy next-generation sequencing (NGS) data deconvolution to interpret your screening results and allows to perform binary and multiparametric functional assays. Furthermore, you can analyze your CRISPR edits in seconds using Sanger sequencing with our CE tool. Our unique multi-guide design ensures you can genotype with one single reaction.

Figure 1. EditCo Arrayed Libraries Workflow. Incorporating smart guide design with automation coupled with our high-quality control, we plate only the best-quality sgRNA into each well with high precision enabling you to reliably knock out your targets in your screens with confidence, we guarantee it (excluding non-essential genes). Due to our optimized workflow, we can deliver your Arrayed CRISPR gRNA Libraries in as few as 7 days at unrivaled speed.

Powered by our unique multi-guide design, we offer both Whole Human and Whole Mouse Genome libraries. Our Whole Genome Libraries ensure the highest editing efficiencies, decreasing false negatives. Our libraries are also delivered ready-to-transfect, minimizing laborious preparation across over 20,000 gene targets.

Our Whole Human and Whole Mouse Genome libraries enable you to get the most of your target identification screen, but if you are more interested in studying your specific gene set or a specific pathway, check out our User-defined or Pathway Libraries.