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Genetic toolkits for engineering Rhodococcus species with versatile applications.

Youxiang Liang1, Huimin Yu2

  • 1Department of Chemical Engineering, Tsinghua University, Beijing 100084, China; Key Laboratory of Industrial Biocatalysis (Tsinghua University), the Ministry of Education, Beijing 100084, China.

Biotechnology Advances
|April 6, 2021
PubMed
Summary

Genetic engineering tools for Rhodococcus bacteria are expanding, addressing challenges like low efficiency. This review details available molecular tools for enhanced biocatalysis and bioremediation applications.

Keywords:
CRISPR/Cas9DNA transformationGenetic toolkitsGenome editingPromotersRhodococcusShuttle vectors

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Area of Science:

  • Microbiology
  • Synthetic Biology
  • Biotechnology

Background:

  • Rhodococcus species are non-model bacteria with significant industrial applications in biocatalysis, bioremediation, and biomass conversion.
  • Engineering Rhodococcus is difficult due to limited molecular tools, high GC content, and low transformation efficiencies.

Purpose of the Study:

  • To comprehensively review the genetic elements and engineering tools developed for Rhodococcus species.
  • To highlight advancements in gene transfer, transformation efficiency improvement, and genome editing techniques.

Main Methods:

  • Review of recently developed genetic elements: shuttle vectors, promoters, antibiotic markers, ribosome binding sites, and reporter genes.
  • Summary of gene transfer techniques and strategies to enhance transformation efficiency.
  • Overview of genome editing tools: transposition, homologous recombination, recombineering, and CRISPR/Cas9.

Main Results:

  • A growing toolkit of genetic elements and engineering strategies is now available for Rhodococcus.
  • Recent advancements have addressed challenges in Rhodococcus genetic manipulation, improving accessibility for researchers.
  • Diverse tools for random and precise genome editing are established.

Conclusions:

  • The development of genetic tools is crucial for unlocking the full potential of Rhodococcus for bioproduction.
  • Future trends point towards integrating synthetic and systems biology approaches for advanced Rhodococcus engineering.
  • Expect optimization of multiplex genome editing, dynamic regulation, and metabolic models for Rhodococcus.