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Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
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Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
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Ribosomes01:27

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Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
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RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
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Orthogonal Ribosome Biofirewall.

Bin Jia1,2, Hao Qi1,2, Bing-Zhi Li1,2

  • 1Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin 300072, PR China.

ACS Synthetic Biology
|August 8, 2017
PubMed
Summary
This summary is machine-generated.

We developed an orthogonal ribosome biofirewall for biocontainment. This system uses genetic circuits to control genetically modified organisms, enhancing biosafety in research and industry.

Keywords:
biocontainmentbiofirewallbiosafetygenetically modified organismsorthogonal ribosomesynthetic biology

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Area of Science:

  • Synthetic Biology
  • Genetic Engineering
  • Biosafety

Background:

  • Biocontainment is essential for preventing the escape of genetically modified organisms (GMOs) into ecosystems.
  • Existing biocontainment methods require improvement for robust control.

Purpose of the Study:

  • To develop and validate a novel orthogonal ribosome biofirewall system for enhanced biocontainment.
  • To demonstrate the flexibility and efficacy of the biofirewall in controlling gene expression.

Main Methods:

  • Designed an activation circuit as a genetic AND gate using orthogonal ribosomes and environmental signals.
  • Engineered a degradation circuit as a genetic NOT gate producing I-SceI endonuclease.
  • Integrated the activation circuit into plasmid-based and genome-based genetic systems.
  • Validated the degradation circuit's ability to reduce target gene and plasmid expression.

Main Results:

  • The activation circuit enabled tightly regulated encryption of the deoxychromoviridans pathway and lacZ, reducing expression to 7.3% and 7.8%.
  • The degradation circuit effectively reduced target plasmid and orthogonal rRNA (O-rRNA) expression to 0.8% in lab medium and 0.76% in soil medium.
  • Demonstrated successful incorporation into genetic circuits and metabolic pathways.

Conclusions:

  • The orthogonal ribosome biofirewall is a versatile platform for biocontainment.
  • This system offers significant potential for biosafety applications in research and the biotechnology industry.