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Related Concept Videos

Initiation of Translation02:33

Initiation of Translation

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Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
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Leaky Scanning02:28

Leaky Scanning

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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
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Regulated mRNA Transport02:22

Regulated mRNA Transport

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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

Types of RNA

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Overview
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.
RNA...
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Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

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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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Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

10.6K
The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
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Related Experiment Video

Updated: Jul 5, 2025

The Encapsulation of Cell-free Transcription and Translation Machinery in Vesicles for the Construction of Cellular Mimics
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The Encapsulation of Cell-free Transcription and Translation Machinery in Vesicles for the Construction of Cellular Mimics

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Cell-Free Expressed Membraneless Organelles Inhibit Translation in Synthetic Cells.

Abbey O Robinson1, Jessica Lee2, Anders Cameron1

  • 1Department of Genetics, Cell Biology and Development, University of Minnesota, 420 SE Washington Ave., Minneapolis, Minnesota 55455, United States.

ACS Biomaterials Science & Engineering
|January 16, 2024
PubMed
Summary

Researchers created artificial membrane-less organelles in synthetic cells. These compartments sequester RNA, effectively inhibiting protein expression and offering insights into natural organelle function.

Keywords:
cell-free expressioncoacervatesmembraneless organellessynthetic biologysynthetic cellstranslation

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Area of Science:

  • Synthetic biology
  • Cell biology
  • Biophysics

Background:

  • Cellular compartments create specialized microenvironments for efficient reactions.
  • Organelles can be membrane-bound or formed via liquid-liquid phase separation (e.g., P-granules, nucleoli).
  • Synthetic minimal cells are valuable tools for studying natural cellular processes like organelle formation.

Purpose of the Study:

  • To engineer artificial membrane-less organelles within synthetic cells.
  • To investigate the function of these artificial organelles in modulating cellular processes.
  • To advance the understanding of natural organelle evolution and synthetic cell complexity.

Main Methods:

  • Expression of RGG-GFP-RGG, a phase-separating protein from *Caenorhabditis elegans* P-granules, using cell-free transcription and translation.
  • Formation of artificial membraneless organelles within synthetic cells.
  • Analysis of RNA sequestration and protein expression inhibition within these engineered compartments.

Main Results:

  • Successfully formed artificial membraneless organelles in synthetic cells.
  • Demonstrated that these organelles sequester RNA, leading to reduced protein expression.
  • Established these compartments as a tool to inhibit protein synthesis in synthetic systems.

Conclusions:

  • Artificial membrane-less organelles can be engineered in synthetic cells to create functional microenvironments.
  • These engineered compartments effectively inhibit protein expression by sequestering RNA.
  • The study contributes to understanding natural organelle function and developing advanced synthetic lifelike systems.