Heterologous Expression and Purification of a CRISPR-Cas9-Based Adenine Base Editor

Seu Na Lee, Hong Su Jang, Jae Sung Woo

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

CRISPR-cas9-guided adenine base editors (ABEs) site-specifically convert the A-T base pair to G-C base pair in genomic DNA. The intracellular delivery of ABE proteins preassembled with guide RNAs (gRNAs) has shown greatly reduced off-target effects compared with that of plasmids or viral vectors containing ABE and gRNA-encoding sequences. For efficient gene editing by the ribonucleoprotein delivery method, the ABE-gRNA complexes need to be prepared in high purity and quantity. Here we describe the expression and purification procedure of ABEmax, one of high-efficiency ABE versions.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages123-133
Number of pages11
DOIs
Publication statusPublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2606
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Funding Information:
This work was supported by the National Research Foundation Grant funded by the Ministry of Science and ICT (NRF-2018R1C1B6004447 to J.-S.W.).

Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

  • Adenine base editor
  • CRISPR-cas9
  • Gene editing
  • Protein purification
  • Ribonucleoprotein

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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