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Function and Application of Type IIS Restriction Enzymes

Learn about the functionality of Type IIS restriction enzymes and their role in molecular cloning, RNA therapeutic and vaccine development.

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Type IIS restriction enzymes are useful for molecular cloning applications, as well as RNA therapeutic and vaccine development workflows. Unlike the more common Type IIP restriction enzymes, such as EcoRI, which cleave symmetric or palindromic sites and typically cleave within the recognition sequence, Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence, often leaving overhang sequences from 1 to 5 basepairs. Type IIS enzymes are useful for many applications, including Golden Gate Assembly. Type IIS enzymes that generate 3- to 5-basepair overhangs, such as PaqCI, BsaI-HFv2 and BsmBI-v2, are commonly used in Golden Gate Assembly. In this method, restriction enzyme sites are removed upon cleavage, resulting in seamless assembly of DNA fragments. New England Biolabs has focused research on the end-joining fidelity of these overhang sequences, leading to the development of the NEBridge Ligase Fidelity Tools. These tools aid in the design of high-fidelity assemblies. Type IIS enzymes are also used in the linearization of plasmids for mRNA therapeutics and vaccine development. For example, BspQI, which is now available in GMP-grade. To learn more about Type IIS restriction enzymes and the applications they are used for, please visit these resources.

Function and Application of Type IIS Restriction Enzymes

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