Related Experiment Video
Updated: Sep 4, 2025
09:51
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
34.3K
Pioneer Factor Improves CRISPR-Based C-To-G and C-To-T Base Editing
Chao Yang1,2, Xingxiao Dong3, Zhenzhen Ma4
1Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|July 21, 2022
Summary
Engineered base editors (BEs) fused with SOX2 pioneer factor significantly enhance gene editing efficiency and broaden the editing window in eukaryotes. This novel approach improves chromatin accessibility for precise genetic modifications.
Area of Science:
- * Molecular Biology
- * Epigenetics
- * Gene Editing Technologies
Background:
- * Base editing efficiency in eukaryotes is influenced by chromatin accessibility.
- * Limited research exists on how specific chromatin factors impact base editor performance.
- * Pioneer factors, like SOX2, are known to remodel chromatin structure.
Purpose of the Study:
- * To investigate the role of pioneer factors in enhancing base editing efficiency and window.
- * To engineer novel base editors fused with SOX2 or its functional domains.
- * To assess the impact of these engineered base editors on chromatin accessibility and editing outcomes.
Main Methods:
- * Construction of base editors (BEs) fused with SOX2 and its functional domains (e.g., SoxN-GBE, SoxM-GBE/CBE, SadN-GBE, SadM-CBE).
- * Evaluation of editing efficiency and editing window at various genomic loci.
- * Chromatin accessibility analysis using DNase I assays.
- * Application of engineered base editors for gene editing in the MYC proto-oncogene.
Main Results:
- * SOX2 fusion significantly increased base editing efficiency for both GBE (guanine base editor) and CBE (cytosine base editor).
- * SoxN-GBE enhanced overall editing efficiency, while SoxM-GBE/CBE improved editing at PAM-proximal cytosines.
- * Engineered SadN-GBE and SadM-CBE demonstrated higher editing efficiency and broader editing windows, respectively.
- * DNase I assays confirmed that SOX2 domains enhance editing by increasing chromatin accessibility.
- * SadM-CBE successfully introduced a stop codon in the MYC proto-oncogene at a challenging locus.
Conclusions:
- * Fusion of pioneer factors or their domains with base editors creates a new class of highly efficient gene editing tools.
- * Engineered pioneer-base editors (pioneer-BEs) offer improved editing efficiency and broader editing windows in eukaryotic systems.
- * This strategy provides a powerful platform for precise genome engineering, particularly at difficult-to-edit sites.
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