A One-Step Method for Efficient Purification of Functional Cas9 Protein

BIO-PROTOCOL 2026 Open access

Read full text
Authors: Xinzhi Duan, Zhengrong Zhou, Aihua Mao
DOI: 10.21769/bioprotoc.5594
Cited by: 0

Abstract

using Ni-NTA chromatography, eliminating the protease cleavage step. • This method yields over 8 mg/L of high purity (>95%), functional Cas9 protein, suitable for direct use in RNP complex assembly. • The purified His-Ub-Cas9 maintains high genome editing activity in vivo, as demonstrated in zebrafish embryos.

Fields of Study & Subjects

CRISPR and Genetic Engineering RNA regulation and disease Protein purification and stability

Publication details

Journal / source
BIO-PROTOCOL
Volume
16
Issue
1390
Pages
e5594–e5594
Publication date
1 January 2026
Language
EN