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Benchling vs snapgene12/5/2023 Golden GATEway cloning kits combine the efficiency of Golden Gate assembly with the sequence-independent approach of Multisite Gateway™ cloning.Īs described by Stephan Kirchmaier and colleagues in PLOS One, fragments of interest are cloned into one of eight Golden Gate entry vectors. Once bound to DNA, the fused nuclease creates a targeted double-strand break.Īn alternative to CRISPR, TALENs are useful genome engineering tools that can be used to create gene knockouts or introduce specific DNA sequence modifications. TAL effectors contain a customizable array of amino acid repeats that bind to desired regions of DNA. FokI), which is achieved through Golden Gate assembly. TALENs are fusions of transcription activator-like (TAL) effectors to a Type IIS restriction endonuclease (e.g. Thomas Cermak and colleagues first described the design and assembly of custom TALEN constructs in 2011. There are various pre-designed kits available that utilize Golden Gate technology, such as transcription activator-like effector nucleases (TALENs), Golden GATEway and modular cloning (MoClo). If the Type IIS recognition site is in a coding sequence, the mutation(s) you introduce must be silent. Unwanted cleavage sites can be removed with site-directed mutagenesis.
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