ArticleNucleic acids research2024
Multiplexed in-situ mutagenesis driven by a dCas12a-based dual-function base editor.
Article in Nucleic acids research, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 20 citations in OpenAlex.
- Leveraging the bacteria for enhanced cancer immunotherapy: from a perspective of synthetic biology.Cancer biology & therapy · 2026Review
- Engineering of multiple modules to enhance lignocellulose degradation ability inSynthetic and systems biotechnology · 2026Article
- Development of Bacillus subtilis cell factories for nutraceuticals production.World journal of microbiology & biotechnology · 2026Review
- 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
- Precision hydrolysis: tailored yeast processing enzymes for yeast-based products.Applied microbiology and biotechnology · 2026Review
- Whole-cell and cell-free biosensor-driven metabolic engineering.Current opinion in biotechnology · 2025Review
- CRISPR-DNA Polymerase Assisted Targeted Mutagenesis for Regulable Laboratory Evolution.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 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
- Precision multiplexed base editing in human cells using Cas12a-derived base editors.Nature communications · 2025Article
- A cross-species inducible system for enhanced protein expression and multiplexed metabolic pathway fine-tuning in bacteria.Nucleic acids research · 2025Article
- CRISPR/Cas13X-assisted programmable and multiplexed translation regulation for controlled biosynthesis.Nucleic acids research · 2025Article
Corrections and comments
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Authors and funding
12 authors at 2 institutions in 2 countries.
Funding
Abstract
Mutagenesis driving genetic diversity is vital for understanding and engineering biological systems. However, the lack of effective methods to generate in-situ mutagenesis in multiple genomic loci combinatorially limits the study of complex biological functions. Here, we design and construct MultiduBE, a dCas12a-based multiplexed dual-function base editor, in an all-in-one plasmid for performing combinatorial in-situ mutagenesis. Two synthetic effectors, duBE-1a and duBE-2b, are created by amalgamating the functionalities of cytosine deaminase (from hAPOBEC3A or hAID*Δ ), adenine deaminase (from TadA9), and crRNA array processing (from dCas12a). Furthermore, introducing the synthetic separator Sp4 minimizes interference in the crRNA array, thereby facilitating multiplexed in-situ mutagenesis in both Escherichia coli and Bacillus subtilis. Guided by the corresponding crRNA arrays, MultiduBE is successfully employed for cell physiology reprogramming and metabolic regulation. A novel mutation conferring streptomycin resistance has been identified in B. subtilis and incorporated into the mutant strains with multiple antibiotic resistance. Moreover, surfactin and riboflavin titers of the combinatorially mutant strains improved by 42% and 15-fold, respectively, compared with the control strains with single gene mutation. Overall, MultiduBE provides a convenient and efficient way to perform multiplexed in-situ mutagenesis.
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