Guidelines for Selection of Gene Editing Tools: From CRISPR to Base Editing

Main Types of Common Gene Editing Tools

Currently common types of gene editing tools include CRISPR-Cas systems, TALENs, ZFNs, and base editing and pioneer editing developed on this basis.The CRISPR-Cas system relies on guided RNA to recognize target sequences, which is relatively simple to operate and suitable for multi-site editing. TALEN and ZFN are recognized through protein-DNA binding domains, and the modular design is relatively mature. Base editing can achieve single-base conversion without double-strand breaks, which is commonly used for point mutation model construction.Different routes have different identification methods, editing windows, and delivery requirements, and need to be evaluated in conjunction with specific experimental objectives.

CRISPR-Cas: RNA guidance, flexible design, attention to Pam sequence restrictions;

TALEN/ZFN: protein guidance, long recognition length, relatively high module construction cost;

Base editing: suitable for single-base substitution, with low dependence on double-strand breaks;

Pilot editing: Insertion, deletion and replacement can be achieved, and the design complexity is high.

Key considerations when choosing a gene editing tool

When selecting tools, it is generally necessary to pay attention to target gene characteristics, cell types, delivery methods, and downstream screening strategies.For primary cells or refractory systems, delivery efficiency can be a limiting factor, taking into account common modes such as viral vectors, electrotransfer, or lipid nanoparticles.Editing efficiency and off-target risk need to be evaluated in parallel, and it is generally recommended to set a negative control and a known site control, combined with