CRISPR Reagents: Cas9 Nucleases, gRNA Control Kits, CRISPR Transfection Reagents, and more

CRISPR controls are an essential component of gene-editing experiment workflows, ensuring accuracy, efficiency, and reproducibility. When building an assay, these controls help optimize workflows across different cell lines and establish a baseline for comparison. During the screening process, they distinguish gene-editing effects from experimental artifacts and assess editing efficiency. Finally, CRISPR controls play a crucial role in optimizing, validating results and troubleshooting when needed. Evaluating whether and how well genome editing worked is just as critical as the outcome itself.

Positive controls

CRISPR positive controls are necessary to validate and monitor the efficiency of the CRISPR reagents being used in any gene editing experiment. These positive controls are validated sgRNA sequences that have demonstrated high editing efficiency across different cell types and are available in both XDel design and single-guide format.

These controls are ideal for building and optimizing your assay, optimizing your experimental conditions, monitoring the gene editing efficiency across different cell lines or workflows, and troubleshooting. They can be used in any cell type, including immortalized, iPSC, and primary cells.

* Each cell line was genetically modified using EditCo’s proprietary process under optimized editing conditions. Editing efficiency was evaluated 72 hours post-nucleofection. The targeted genomic region was amplified via PCR, sequenced using Sanger sequencing, and analyzed with the ICE analysis tool to determine knockout (KO) levels. To assess the impact of CRISPR editing on cell growth, total cell numbers were measured using a fluorescence-based cell counting assay 72 hours post-nucleofection. Hoechst staining was used to label all cells, while dead cells were identified via propidium iodide staining. The number of live cells was determined by subtracting dead cells from the total count.

** Data from a representative control is highlighted below.

Figure 1. Positive controls provide high editing efficiency without affecting cell viability. A) Knockout score and B) Cell viability for representative positive control, IFNGR, versus negative control, NTC3. 

Negative controls

A negative control in a CRISPR-Cas9 experiment is used to ensure that observed effects are due to the intended genetic perturbation rather than artifacts from the experimental workflow (such as off-target effects, cell stress, or other non-specific influences). These non-targeting gRNA sequences are designed to not target any genomic location in the human & mouse species and therefore no editing event occurs with SpCas9. These controls are validated sequences that have been shown to have no effect on cell viability across different cell types and are available in a single-guide format.

These controls are essential in gene-editing experiments to compare the phenotype of cells treated with a gene-targeting guide RNA to those treated with a non-targeting guide RNA. They are commonly included in CRISPR screens to help identify potential false positives.

These kits include positive controls, available in either XDel or single-guide format, that are useful for optimizing transfection conditions or gaining familiarity with CRISPR/Cas9 technology before targeting your gene of interest. Each kit contains everything needed for a complete CRISPR experiment, including controls, SpCas9 protein, and primers.

The Transfection Optimization Kit (TOK) is designed to be used alongside the Gene Knockout Kit, as both leverage the XDel strategy for efficient gene editing.

* Each cell line was genetically modified using EditCo’s proprietary process under optimized editing conditions. Editing efficiency was evaluated 72 hours post-nucleofection. The targeted genomic region was amplified via PCR, sequenced using Sanger sequencing, and analyzed with the ICE analysis tool to determine knockout (KO) levels. To assess the impact of CRISPR editing on cell growth, total cell numbers were measured using a fluorescence-based cell counting assay 72 hours post-nucleofection. Hoechst staining was used to label all cells, while dead cells were identified via propidium iodide staining. The number of live cells was determined by subtracting dead cells from the total count. 

EditCo offers high-quality Streptococcus pyogenes Cas9 protein with two nuclear localization signals (2NLS) for enhanced nuclear delivery. This SpCas9 protein has been validated and demonstrates high editing efficiency across various loci in both in vitro and ex vivo experiments. Cas9 in protein form is ideal for RNP-based CRISPR experiments, enabling rapid and transient genome editing with minimal off-target effects. Delivered directly as a ribonucleoprotein (RNP) complex with guide RNA, it eliminates the need for cellular transcription or translation, providing fast action and greater control over editing outcomes. The protein is also compatible with multiple transfection methods, including electroporation and lipofection.

* HEK293T cells were genetically edited using EditCo’s proprietary process under optimized conditions to target an endogenous genomic site. Editing efficiency was assessed 72 hours post-nucleofection. The targeted genomic region was amplified via polymerase chain reaction (PCR), sequenced using Sanger sequencing, and analyzed with the ICE (Inference of CRISPR Edits) tool to quantify knockout (KO) levels.

We offer Tris-EDTA Buffer and Nuclease-free Water that can be used to resuspend or dilute your gRNA libraries.

These controls can be purchased as kits, individually along with SpCas9 protein through our add-ons page or can be added in your custom arrayed library.