What is the best way to deliver the ZFN and the repair template into the cells?
The ZFN itself can be delivered as a plasmid or it can be transcribed into an mRNA transcript for delivery. We have found both plasmid and mRNA to be effective depending on cell type and, therefore, provide customers with ZFNs in both plasmid and mRNA form to test in their own cell lines. Repair templates are typically delivered into the cell as linear DNA. Regardless of form, mRNA, DNA, or plasmid, we have found that the use of nucleofection or electroporation generally produce the highest DNA cleavage efficiencies. Due to this increase in efficiency, we recommend using an electroporation or nucleofection delivery method if possible. Lipid-based transfection also works in many cases, but may result in lower efficiencies.
