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Related Concept Videos

Translational Regulation01:29

Translational Regulation

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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
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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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In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing...
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Types of RNA01:23

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Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
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Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
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Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
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RNA-responsive elements for eukaryotic translational control.

Evan M Zhao1, Angelo S Mao1,2, Helena de Puig1,2

  • 1Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.

Nature Biotechnology
|October 29, 2021
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Summary
This summary is machine-generated.

Scientists developed eukaryotic toehold switches (eToeholds) to precisely control gene translation in cells. This new riboregulator technology significantly enhances transgene expression and enables specific RNA detection for various applications.

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Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Biotechnology

Background:

  • Controlling gene translation in eukaryotic cells is crucial for biotechnological advancements.
  • Existing methods for RNA-based gene control suffer from limited expression fold changes and large trigger RNA (trRNA) sizes.

Purpose of the Study:

  • To introduce and optimize eukaryotic toehold switches (eToeholds) as a novel class of modular riboregulators for controlling gene expression.
  • To achieve high-fold induction of transgene expression using eToeholds in mammalian cells.
  • To demonstrate the capability of eToeholds to act as RNA sensors for discriminating cellular states.

Main Methods:

  • Designed eToeholds incorporating internal ribosome entry site sequences that form inhibitory loops.
  • Optimized RNA annealing between eToeholds and specific trigger RNAs (trRNAs).
  • Validated eToehold performance in mammalian cells, measuring transgene expression levels.

Main Results:

  • Achieved up to 16-fold induction of transgene expression through optimized eToehold-trRNA interactions.
  • Demonstrated that eToeholds effectively disrupt inhibitory loops upon trRNA binding, enabling translation.
  • Showcased eToeholds' ability to differentiate between viral infection status, gene expression presence/absence, and cell types based on RNA transcripts.

Conclusions:

  • Eukaryotic toehold switches (eToeholds) represent a powerful and modular tool for precise control of gene translation in eukaryotic systems.
  • Optimized eToeholds offer significant improvements in transgene expression fold-change compared to previous methods.
  • eToeholds function as effective RNA biosensors, enabling the detection of specific RNA transcripts for cellular state monitoring.