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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

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Updated: Jan 16, 2026

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Published on: February 28, 2021

MicroDFBEST: A dCas12b-derived dual-function base editor with programmable editing characteristics for microbial

Wen-Liang Hao¹, De-Zhi Geng², Yu-Feng Liu³

¹Key Laboratory of Industrial Biotechnology (Ministry of Education), School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China.

Synthetic and Systems Biotechnology

|October 6, 2025

Summary

We developed MicroDFBEST, a dual-function base editor for microbes, enabling simultaneous C-to-T and A-to-G genome editing. This versatile tool expands microbial gene editing capabilities for synthetic biology applications.

Keywords:

CRISPR/Cas12b Dual-function base editor Genetic diversity In situ evolution Targeted mutagenesis

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Published on: January 25, 2019

Area of Science:

Microbial biotechnology
Synthetic biology
Genome editing

Background:

Existing base editors (BEs) have limitations in microbial applications, including restricted mutagenesis and narrow editing windows.
Developing novel tools is crucial to enhance genome editing precision and versatility in microorganisms.

Purpose of the Study:

To engineer a novel dual-function base editor (DFBE) for microbes, named MicroDFBEST.
To expand the capabilities of base editing in microbial systems for diverse synthetic biology applications.

Main Methods:

Fusing high-activity deaminases (evoCDA1 and TadA9) with a nuclease-deficient Cas12b (dBhCas12b).
Utilizing MicroDFBEST for simultaneous C-to-T and A-to-G editing within a 26-33 nt window.
Adjusting editing characteristics by modifying fusion protein expression and editing generations.

Main Results:

Achieved the broadest editing window reported for microbial DFBEs.
Demonstrated flexible gene expression modulation through promoter diversification (P_ylbP).
Enabled targeted protein evolution via mutational hotspot scanning in native genomic contexts.

Conclusions:

MicroDFBEST provides a versatile platform for in situ gene regulation, such as activating biosynthetic gene clusters.
The system facilitates protein evolution, including chassis optimization, with broad synthetic biology utility.