Methods – Rapid Single-Pot Assembly of Modular Chromatin Proteins for Epigenetic Engineering

Posted on December 6, 2022 Updated on April 21, 2023

Rapid Single-Pot Assembly of Modular Chromatin Proteins for Epigenetic Engineering
Haynes KA, Priode JH. (2023) Methods Mol Biol. 2599:191-214.
PMID: 36427151

Much of epigenetic engineering relies on the assembly of multifunctional fusion proteins. We developed a set of cloning vectors and a protocol for one-step “Golden Gate” construction of recombinant protein-encoding DNA.  Standard 2-amino acid linkers allow flexible assembly of any combination of up to four protein modules, eliminating the need to design different compatible Golden Gate overhangs to ligate different modules. The five cloning vectors described in this protocol are available at Addgene: Karmella Haynes Lab Plasmids.

Related:

• Addgene. Hot Plasmids and Viral Preps – March 2021. https://blog.addgene.org/hot-plasmids-and-viral-preps-march-2021
• Rapid Single-Pot Assembly of Modular Chromatin Proteins for Epigenetic Engineering
  Priode JH, Haynes KA. (2021) Protocols.io. https://dx.doi.org/10.17504/protocols.io.brgcm3sw