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Related Concept Videos

Leaky Scanning02:28

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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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Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
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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.
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Directionality of Nuclear Transport01:42

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Ras-related nuclear protein or Ran is a small G protein that cycles between its GTP and GDP bound states. Ran specific regulators, a Ran GTPase Activating Protein or RanGAP present in the cytosol and a Ran guanine nucleotide exchange factor or RanGEF present inside the nucleus regulate GTP/GDP exchange. A high concentration of GTP inside the cells, in addition to this asymmetric distribution of  Ran-specific regulators, leads to a higher RanGTP concentration inside the nucleus. This...
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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.
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RACE - Rapid Amplification of cDNA Ends02:35

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Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...
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Updated: Oct 3, 2025

Measuring RAN Peptide Toxicity in C. elegans
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Measuring Repeat-Associated Non-AUG (RAN) Translation.

Shaopeng Wang1, Shuying Sun2

  • 1Department of Pathology, Physiology, Brain Science Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 16, 2022
PubMed
Summary
This summary is machine-generated.

Short nucleotide repeat expansions cause over 50 diseases. Researchers developed a dual-luciferase reporter system to measure repeat-associated non-AUG (RAN) translation, aiding the study of diseases like ALS and FTD.

Keywords:
Cap-independentIntegrated stress responseRAN translationRepeat expansioneIF2α phosphorylation

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Expansions of short nucleotide repeats are implicated in over 50 neurological and neuromuscular diseases.
  • Repeat expansion-containing RNAs can produce toxic repeat proteins via repeat-associated non-AUG (RAN) translation.
  • Understanding RAN translation mechanisms is crucial for elucidating disease pathogenesis.

Purpose of the Study:

  • To describe a dual-luciferase reporter system for quantitatively measuring RAN translation.
  • To provide a platform for examining candidate genes and pathways involved in RAN translation.
  • To screen for modifiers of this non-canonical translation pathway.

Main Methods:

  • Development and application of a dual-luciferase reporter system.
  • Quantitative measurement of RAN translation efficiency.
  • Utilizing the C9ORF72 GGGGCC repeat expansion as a model system.

Main Results:

  • The dual-luciferase reporter system enables quantitative assessment of RAN translation.
  • This system can be used to study the C9ORF72 GGGGCC repeat expansion, a key factor in ALS and FTD.
  • The described method facilitates the investigation of factors influencing RAN translation.

Conclusions:

  • The dual-luciferase reporter system is a valuable tool for studying RAN translation.
  • This system aids in understanding the molecular mechanisms underlying repeat expansion diseases.
  • It provides a foundation for identifying therapeutic targets for conditions like ALS and FTD.