Sometimes dead cells are a good thing! Universal cell death controls for CRISPR-Cas9 optimization.
Appropriate controls are essential for good experimental design. Since gene editing requires delivery of the CRISPR-Cas9 components to generate double-strand breaks as well as imperfect repair by endogenous mechanisms of the cell, effective negative and positive controls are critical for achieving efficient gene knockout and interpreting results. Non-targeting controls enable assessment of potential non-specific effects, while positive controls can be used to assess the efficiency of gene editing and serve as an indicator of good transfection in ongoing experiments.
Delivery of CRISPR-Cas9 components to cells is relatively fast, occurring within hours. However, the development of a pronounced phenotype due to target gene knockout can take days. It is beneficial to a researcher to have a degree of certainty – earlier in the process – that the experiment has been successful, at least from the standpoint of delivery efficiency. This is particularly useful if the measured phenotype has a longer incubation time. While initial experimental optimization should be performed using a target-specific positive control in which editing efficiency can be quantitatively assessed, during many experiments a rapid indication of delivery efficiency can save resources, labor, reagents, and time, that may otherwise be wasted on performing costly and time-consuming phenotypic assays on failed experiments or interpretation of weak results due to poor delivery.
Edit-R Lethal crRNA controls are universal positive controls based on cell death, such that cell viability can be correlated to the efficiency of the delivery of CRISPR-Cas9 components to the cells. They provide quick, visual, and quantifiable indication of transfection efficiency.
Species-specific crRNAs targeting well-characterized genes, as well as mismatch detection assay primers, to determine the effectiveness of your gene editing conditions for maximal efficiency.
A convenient table to help you choose effective controls for your gene editing experiment
Whether your goal is gene knockout or creating a knock-in, this guide will assist you in selecting the right Edit-R tools for your genome engineering application.