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

Seu Na Lee; Hong Su Jang; Jae Sung Woo

doi:10.1007/978-1-0716-2879-9_10

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

Seu Na Lee, Hong Su Jang, Jae Sung Woo

Department of Life Sciences

Research output: Chapter in Book/Report/Conference proceeding › Chapter

1 Citation (Scopus)

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 language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	123-133
Number of pages	11
DOIs	https://doi.org/10.1007/978-1-0716-2879-9_10
Publication status	Published - 2023

Publication series

Name	Methods in Molecular Biology
Volume	2606
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

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

Access to Document

10.1007/978-1-0716-2879-9_10

Cite this

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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.",

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AB - 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.

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KW - Protein purification

KW - Ribonucleoprotein

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

Abstract

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Bibliographical note

Keywords

ASJC Scopus subject areas

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