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Related Concept Videos

Ribozymes02:47

Ribozymes

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The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can...
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Riboswitches01:56

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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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Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
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Regulation of Expression at Multiple Steps01:23

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The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
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What is Gene Expression?01:36

What is Gene Expression?

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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
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Tunable self-cleaving ribozymes for modulating gene expression in eukaryotic systems.

Thomas Jacobsen1, Gloria Yi1, Hadel Al Asafen1

  • 1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, United States of America.

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|May 1, 2020
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Summary

Scientists developed new genetic tools for synthetic biology by combining ribozymes and competing sequences to regulate gene expression in eukaryotes. These tools successfully modulated GFP expression in mammalian cells and Drosophila embryos.

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Area of Science:

  • Synthetic biology
  • Molecular biology
  • Genetics

Background:

  • Genetic tools are crucial for advancing synthetic biology and regulating gene expression.
  • Existing gene regulatory tools for eukaryotic systems lag behind those for single-celled organisms.

Purpose of the Study:

  • To engineer novel genetic regulatory tools for eukaryotic systems.
  • To address the limitations of current gene expression control mechanisms in eukaryotes.

Main Methods:

  • Engineered gene regulatory tools by combining self-cleaving ribozymes with upstream competing sequences.
  • Designed competing sequences to disrupt ribozyme self-cleavage.
  • Tested tool functionality in mammalian cells and Drosophila embryos by modulating GFP expression.

Main Results:

  • Successfully modulated Green Fluorescent Protein (GFP) expression in mammalian cells.
  • Demonstrated the feasibility of these tools in Drosophila embryos.
  • Gene expression reduction varied based on the location (5' vs. 3' untranslated region) and type of competing sequence used.

Conclusions:

  • Developed a novel set of genetic tools for precise gene expression regulation in eukaryotes.
  • These tools offer a versatile method for tuning gene expression across diverse eukaryotic systems.
  • The findings expand the toolbox for synthetic biology applications in complex organisms.