In CRISPR-Cas9 gene editing experiments, Cas9 nuclease binds to gRNA and induces a double-stranded break at a specific genomic target sequence. For the Cas9 enzyme to function, this target sequence must have a proto-spacer adjacent motif (PAM) site in the vicinity of the desired break. Cas9 nuclease is available in several formats to fit your experiment design and preferred workflow, allowing for precise DNA cleavage and gene knockout.
|Designed for||Most CRISPR research and pre-clinical applications||Stable cell line creation|
|Key limitations||None||Requires translation||Potential for cellular response to viral transduction|
|Recommended delivery options||Lipofectamine CRISPRMAX Transfection Reagent||Lipofectamine MessengerMAX Transfection Reagent||Lipofectamine 3000 Transfection Reagent||Lentiviral transduction|
|Learn more||Cas9 protein options||Cas9 mRNA options||Cas9 lentivirus options|
In contrast to the plasmid and mRNA formats, transfection of our TrueCut Cas9 protein complexed with our TrueGuide synthetic gRNA bypasses transcription and translation, helping to sharply increase editing efficiency by the Cas9 enzyme in your experiments. Furthermore, while CRISPR plasmids remain in the cell for more than 72 hours and may exacerbate off-target cleavages, TrueCut Cas9 proteins are cleared from the cell within 24 hours, minimizing off-target effects.
Figure 1. Comparing CRISPR-Cas9 gene editing formats. Cas9 and guide RNA can be delivered to cells as DNA, RNA or pre-formed ribonucleoprotein complexes (RNPs) formats. When using a plasmid DNA vector (upper left), the Cas9 gene must first be transcribed in the nucleus, exported to the cytoplasm, and translated, while an mRNA/gRNA mix (bottom) must be translated. A reagent-protein complex (upper right), like a TrueCut Cas9 protein coupled with our a TrueGuide synthetic gRNA, bypasses these preliminary steps (i.e., transcription and translation) for simpler and more efficient gene editing.
For Research Use Only. Not for use in diagnostic procedures.