Advances and applications of gene editing and transcriptional regulation in electroactive microorganisms

CHEN Yaru; CAO Yingxiu; SONG Hao

doi:10.12211/2096-8280.2023-056

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Advances and applications of gene editing and transcriptional regulation in electroactive microorganisms

Invited Review | 更新时间：2025-03-20

- Advances and applications of gene editing and transcriptional regulation in electroactive microorganisms
- Synthetic Biology Journal Vol. 4, Issue 6, Pages: 1281-1299(2023)
- 作者机构：
  
  1.天津大学，化工学院，天津 300072
  2.天津大学，合成生物学前沿科学中心，系统生物工程教育部重点实验室，天津 300072
- 作者简介：
- 基金信息：
- DOI：10.12211/2096-8280.2023-056
  CLC： Q812
- Received：19 August 2023，
  
  Revised：2023-09-18，
  
  Published：31 December 2023
- 稿件说明：
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陈雅如，曹英秀，宋浩. 电活性微生物基因编辑与转录调控技术进展与应用［J］. 合成生物学， 2023， 4（6）： 1281-1299

CHEN Yaru， CAO Yingxiu， SONG Hao. Advances and applications of gene editing and transcriptional regulation in electroactive microorganisms［J］. Synthetic Biology Journal， 2023， 4（6）： 1281-1299
陈雅如，曹英秀，宋浩. 电活性微生物基因编辑与转录调控技术进展与应用［J］. 合成生物学， 2023， 4（6）： 1281-1299 DOI： 10.12211/2096-8280.2023-056.

CHEN Yaru， CAO Yingxiu， SONG Hao. Advances and applications of gene editing and transcriptional regulation in electroactive microorganisms［J］. Synthetic Biology Journal， 2023， 4（6）： 1281-1299 DOI： 10.12211/2096-8280.2023-056.

摘要

电活性微生物通过胞外电子传递通路与胞外电子受体/供体进行双向电子交换，产生或吞噬电流。电活性微生物已广泛应用于微生物电化学技术领域，涵盖了元素的生物地球化学循环、环境污染的生物处理与电能生产、生物传感、微生物冶金以及化学品的微生物电合成等多个领域，成为全球环境保护和低碳经济的研究热点。然而，这些微生物在实际应用中仍面临较大局限，如微生物燃料电池的输出功率密度存在一定的上限、微生物电合成技术中的CO

还原速率尚未达到理想水平等。为了克服这些限制性因素，需要通过高效的基因编辑和转录调控策略来改变电活性微生物的遗传特性，提高其双向电子传递效率。本文首先总结了模式电活性微生物（希瓦氏菌和地杆菌）和其他代表性电活性微生物的基因编辑方法和利用CRISPR（clustered regularly interspaced short palindromic repeat）技术实现转录调控的策略。在基因编辑方面，涵盖了（CRISPR辅助的）同源重组、碱基编辑等方法；而在转录调控方面，包括了CRISPR介导的抑制和激活。此外，对于多基因编辑和调控的策略也进行了深入探讨。其次，综述了这些技术在环境、能源领域中的应用，包括微生物燃料电池、污染物生物处理和修复等。最后，讨论了目前电活性微生物工程改造所面临的挑战和未来的发展方向。

Abstract

Electroactive microorganisms (EAMs) engage in bidirectional electron exchange with extracellular electron acceptors/donors through the extracellular electron transfer (EET) pathways

resulting in the generation or consumption of electric current. EAMs have b

een widely applied in many microbial electrochemical technologies

such as biogeochemical cycling of Earth elements

bioremediation of environmental pollutants

electricity production

biosensing

biomining

and microbial electrosynthesis of chemicals

rendering EAMs a focal point in the global pursuit of environmental conservation and low-carbon economy. However

there are still substantial limitations in the practical applications of EAMs. For example

microbial fuel cells encounter a capped upper limit in power density

and the CO

reduction rate in microbial electrosynthesis remains below the desired threshold for practical applications. Overcoming these challenges necessitates enhancement in the bidirectional EET rate of EAMs. Nonetheless

complex phenotypes such as elevated EET efficiency often correlate with the expression of multiple genes. To obtain high-performance EAM strains

a deep comprehension of the genotype-phenotype relationship in EAMs and more nuanced manipulation at the genomic level are imperative. This review provides a comprehensive summary of the latest advances in genome editing and transcriptional regulation in EAMs. The main focus is on the CRISPR (clustered regularly interspaced short palindromic repeat)-based biotechnologies developed in model EAMs

such as

Shewanella

oneidensis

and

Geobacter

sulfurreducens

and a few other representative EAMs. The genome editing techniques to be discussed include (CRISPR-assisted) homologous recombination

CRISPR-associated transposase systems

and base editing. Similarly

transcriptional regulation tools involve CRISPR-based interference (CRISPRi) and activation (CRISPRa) systems. Strategies and advancements related to multiplexed editing and regulation are thoroughly summarized. Subsequently

the review delves into the applications of these technologies in both fundamental and applied scientific domains. On the fundamental science front

efforts are directed toward unve

iling factors related to EET and uncovering hidden genotypes. In the realm of application

green electricity-producing microbial fuel cells

bioremediation of nuclear waste

heavy metals

and azo dyes are discussed. Finally

current challenges and future directions in the genetic engineering of EAMs are discussed.

关键词

Keywords

references

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