Background
The use of CRISPR/Cas9 technologies in generating single-base pair knock-in mutations has recently exploded in the number of
Conclusion
The methods and/or combination of methods outlined in this study can be used to help other researchers with similar goals in single-base pair genome editing.
Results
In this study, we evaluated a number of CRISPR/Cas9 approaches for deriving cell lines with knock-in base pair edits to create a phosphorylation mutation and provide a breakdown of editing efficiencies and suggestions for improvement. Overall, our studies suggest that using pre-formed ribonucleoprotein (RNP) complexes is a reliable editing method to generate homozygous single-base pair mutations. We also show that antibiotic selection coupled homologous recombination is an efficient tool for generating highly specific heterozygous mutations.