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^*

^*Corresponding author for this work

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

2 Citations (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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title = "Heterologous Expression and Purification of a CRISPR-Cas9-Based Adenine Base Editor",

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.",

keywords = "Adenine base editor, CRISPR-cas9, Gene editing, Protein purification, Ribonucleoprotein",

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note = "Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",

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language = "English",

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AU - Lee, Seu Na

AU - Jang, Hong Su

AU - Woo, Jae Sung

PY - 2023

Y1 - 2023

N2 - 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.

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.

KW - Adenine base editor

KW - CRISPR-cas9

KW - Gene editing

KW - Protein purification

KW - Ribonucleoprotein

UR - https://www.scopus.com/pages/publications/85145492403

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

M3 - Chapter

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AN - SCOPUS:85145492403

T3 - Methods in Molecular Biology

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EP - 133

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

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

Abstract

Publication series

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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