ArticleNucleic acids research2021
Development of a base editor for protein evolution via in situ mutation in vivo.
Article in Nucleic acids research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 39 citations in OpenAlex.
- Base Editors for Engineering Industrial Microorganisms: Types, Applications, and Future Perspectives.Biotechnology journal · 2026Review
- MicroDFBEST: A dCas12b-derived dual-function base editor with programmable editing characteristics for microbial genetic engineering.Synthetic and systems biotechnology · 2026Article
- Review
- Advances in gene editing tools for four typical Gram-positive bacteria.Frontiers in microbiology · 2026Review
- Research progress of base editing and prime editing tools based on the CRISPR/Cas system.Molecular therapy. Nucleic acids · 2025Review
- Characterization and engineering of a type IV-A3 CRISPR-Cas system for genome editing in Escherichia coli.Nucleic acids research · 2025Article
- Fueling chromosomal gene diversification and artificial evolution with CRISPR.Genome biology · 2025Review
- T7 RNA polymerase-guided base editor for accelerated continuous evolution inSynthetic and systems biotechnology · 2025Article
- Harnessing an anti-CRISPR protein for powering CRISPR/Cas9-mediated genome editing in undomesticated Bacillus strains.Microbial cell factories · 2025Article
- Development of a CRISPR-based cytosine base editor for restriction-modification system inactivation to enhance transformation efficiency in Vibrio Sp. dhg.Journal of biological engineering · 2025Article
- Engineering of Peptide-Inserted Base Editors with Enhanced Accuracy and Security.Small (Weinheim an der Bergstrasse, Germany) · 2025Article
- A universal and wide-range cytosine base editor via domain-inlaid and fidelity-optimized CRISPR-FrCas9.Nature communications · 2025Article
- A New-Generation Base Editor with an Expanded Editing Window for Microbial Cell Evolution In Vivo Based on CRISPR‒Cas12b Engineering.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Multiplexed in-situ mutagenesis driven by a dCas12a-based dual-function base editor.Nucleic acids research · 2024Article
- Expanding the flexibility of base editing for high-throughput genetic screens in bacteria.Nucleic acids research · 2024Article
- The expanded CRISPR toolbox for constructing microbial cell factories.Trends in biotechnology · 2024Review
- Engineered bacterial orthogonal DNA replication system for continuous evolution.Nature chemical biology · 2023Article
- Construction and application of an efficient dual-base editing platform forChemical science · 2022Article
- CRISPR-Mediated Base Editing: From Precise Point Mutation to Genome-Wide Engineering in Nonmodel Microbes.Biology · 2022Review
- Research progress of pathway and genome evolution in microbes.Synthetic and systems biotechnology · 2022Article
Corrections and comments
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Protein evolution has significantly enhanced the development of life science. However, it is difficult to achieve in vitro evolution of some special proteins because of difficulties with heterologous expression, purification, and function detection. To achieve protein evolution via in situ mutation in vivo, we developed a base editor by fusing nCas with a cytidine deaminase in Bacillus subtilis through genome integration. The base editor introduced a cytidine-to-thymidine mutation of approximately 100% across a 5 nt editable window, which was much higher than those of other base editors. The editable window was expanded to 8 nt by extending the length of sgRNA, and conversion efficiency could be regulated by changing culture conditions, which was suitable for constructing a mutant protein library efficiently in vivo. As proof-of-concept, the Sec-translocase complex and bacitracin-resistance-related protein BceB were successfully evolved in vivo using the base editor. A Sec mutant with 3.6-fold translocation efficiency and the BceB mutants with different sensitivity to bacitracin were obtained. As the construction of the base editor does not rely on any additional or host-dependent factors, such base editors (BEs) may be readily constructed and applicable to a wide range of bacteria for protein evolution via in situ mutation.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.