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Synthetic Biology02:55

Synthetic Biology

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Synthetic biology is an interdisciplinary science that involves using principles from disciplines such as engineering, molecular biology, cell biology, and systems biology. It involves remodeling existing organisms from nature or constructing completely new synthetic organisms for applications such as protein or enzyme production, bioremediation, value-added macromolecule production, and the addition of desirable traits to crops, to name a few.
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Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
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Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
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RoboMoClo: A Robotics-Assisted Modular Cloning Framework for Multiple Gene Assembly in Biofoundry.

Dong Hun Kang1,2, Sung Cheon Ko1,2, Yu Been Heo1,2

  • 1Department of Food Science and Biotechnology, Sungkyunkwan University (SKKU), 2066 Seobu-ro, Jangan-gu, Suwon 16419, Republic of Korea.

ACS Synthetic Biology
|February 15, 2022
PubMed
Summary

Robot-assisted MoClo (RoboMoClo) streamlines synthetic biology by automating DNA assembly in biofoundries. This new system simplifies complex gene expression and pathway construction in bacteria.

Keywords:
Corynebacterium glutamicumMoClobiofoundrygolden gate cloninglycopene

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Bioengineering

Background:

  • Modular cloning (MoClo) systems are crucial for synthetic biology but can be limited in biofoundry automation.
  • Current MoClo workflows require extensive preparation of cloning materials, hindering automation efficiency.

Purpose of the Study:

  • To develop a robot-assisted MoClo (RoboMoClo) framework to accelerate synthetic biology projects within a biofoundry setting.
  • To improve the simplicity and feasibility of multiple DNA assemblies for automated workflows.

Main Methods:

  • Developed RoboMoClo, a hybrid strategy combining hierarchical and iterative gene assembly.
  • Utilized Corynebacterium glutamicum as a model host for RoboMoClo implementation.
  • Constructed biopart libraries (level 0), evaluated gene features (level 1), and performed multiple gene assemblies (level 2).
  • Demonstrated proof-of-concept for biofoundry-assisted lycopene biosynthesis pathway construction (levels 1-3).

Main Results:

  • RoboMoClo enables efficient multiple gene expression and assembly with fewer destination vectors.
  • Investigated transcriptional interference in convergent constructs due to RNA polymerase collision.
  • Successfully demonstrated automated construction of sub-pathway and pathway units for lycopene biosynthesis.

Conclusions:

  • RoboMoClo offers an enhanced MoClo toolkit for laboratory automation in synthetic biology.
  • The framework accelerates synthetic biology projects by simplifying complex DNA assembly processes.
  • RoboMoClo facilitates the construction of engineered metabolic pathways in industrial bacteria.