Compositions and methods for combinatorial assembly and screening for functional site-directednucleotide modifiers

Pollak, Bernardo; Lagos-Susaeta, Diego; Norambuena, Tomás


The present invention discloses a method where a site-directed nuclease (e.g. Cas nuclease) as well as a nucleotide modifying enzyme (e.g. deaminase) are provided as modules of a sequence assembly system, such as a golden gate cloning system, comprising cloning sites and/or regulatory sites. The assembly system provides capacities for generating a polypeptide consisting of at least two standardised modules, each which can be composed of multiple peptide motifs, which optionally can be joined along with other standardised modules for extending the polypeptide's functionality further. New assemblies intended for site-directed nucleotide modification are also provided in this invention.

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US 20230080513 A1