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Non-LTR Retrotransposons

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As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
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LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
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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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In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA...
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Related Experiment Video

Updated: May 5, 2026

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
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LwaCas13a indiscriminately targets the human transcriptome.

Aguilar-Martinez E1, Tonthat G1, Antony Adamson1

  • 1Faculty of Biology. Medicine and Health, University of Manchester, Manchester, Greater Manchester, M13 9PT, UK.

Wellcome Open Research
|May 4, 2026
PubMed
Summary

Leptotrichia wadei Cas13a (LwCas13a) causes cell toxicity and death in mammalian cells due to non-specific RNA targeting. This finding indicates LwCas13a is unsuitable for targeted mRNA downregulation in these systems.

Keywords:
LwaCas13a; down-regulation; off-target; trans-collateral activity; RNA

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • RNA Interference

Background:

  • CRISPR-Cas13a systems offer RNA-targeting gene silencing.
  • Leptotrichia wadei Cas13a (LwCas13a) exhibits RNA degradation in bacteria.
  • Trans-collateral RNA degradation by LwCas13a is reportedly absent in mammalian cells.

Purpose of the Study:

  • To investigate the efficacy of LwCas13a for mRNA downregulation in mammalian cells.
  • To assess the potential off-target effects of LwCas13a in human cell lines.

Main Methods:

  • A doxycycline-inducible system was established to express LwCas13a in HEK293T cells.
  • RNA sequencing (RNA-seq) was employed to analyze LwCas13a off-target activity.

Main Results:

  • LwCas13a activation in HEK293T cells resulted in widespread, non-specific RNA targeting.
  • This non-specific activity led to significant cellular toxicity and cell death.

Conclusions:

  • LwCas13a exhibits detrimental off-target activity in mammalian cells.
  • The observed toxicity renders LwCas13a inappropriate for specific mRNA downregulation applications in human cells.