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Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
Gene Conversion02:08

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
RNA Editing02:23

RNA Editing

RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
Restriction Enzymes01:11

Restriction Enzymes

Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...

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Related Experiment Video

Updated: Jul 18, 2026

Generation of Genome-wide Chromatin Conformation Capture Libraries from Tightly Staged Early Drosophila Embryos
10:35

Generation of Genome-wide Chromatin Conformation Capture Libraries from Tightly Staged Early Drosophila Embryos

Published on: October 3, 2018

Specific cleavage of hyper-edited dsRNAs.

A D Scadden1, C W Smith

  • 1Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1GA, UK

The EMBO Journal
|August 3, 2001
PubMed
Summary

A novel cytoplasmic endonuclease specifically cleaves hyper-edited double-stranded RNA (dsRNA) generated by adenosine deaminases acting on RNA (ADARs). This targeted cleavage offers a pathway for cellular defense against viral RNA.

Area of Science:

  • Molecular Biology
  • Virology
  • RNA Biology

Background:

  • Double-stranded RNA (dsRNA) is a key trigger for cellular antiviral responses.
  • Adenosine deaminases acting on RNA (ADARs) extensively edit dsRNA, converting adenosine to inosine (A-to-I editing).
  • Hyper-edited dsRNA is often associated with viral infections.

Purpose of the Study:

  • To investigate a novel mechanism for the cellular clearance of hyper-edited dsRNA.
  • To identify the specific structural requirements for the cleavage of edited dsRNA.
  • To explore the potential of this mechanism as an antiviral defense strategy.

Main Methods:

  • Characterization of cytoplasmic endonuclease activity.
  • Analysis of cleavage specificity on modified and unmodified dsRNA substrates.

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Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
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Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

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Last Updated: Jul 18, 2026

Generation of Genome-wide Chromatin Conformation Capture Libraries from Tightly Staged Early Drosophila Embryos
10:35

Generation of Genome-wide Chromatin Conformation Capture Libraries from Tightly Staged Early Drosophila Embryos

Published on: October 3, 2018

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
09:12

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation

Published on: September 16, 2019

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
06:51

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

  • Investigation of structural determinants for cleavage, including base pairing (IU/UI vs. GU/UG).
  • Main Results:

    • Discovery of a cytoplasmic endonuclease that specifically targets hyper-edited dsRNA.
    • Cleavage occurs at unique alternating inosine-uridine (IU) and uridine-inosine (UI) base pair sites.
    • Unmodified dsRNA and dsRNA with non-alternating or isomorphic base pairs are resistant to cleavage.

    Conclusions:

    • Hyper-edited dsRNA possesses a unique structure recognized by a specific endonuclease.
    • This cleavage mechanism provides a targeted pathway for the disposal of viral dsRNA.
    • ADAR-mediated hyper-editing followed by endonuclease cleavage represents a novel antiviral defense pathway.