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Related Concept Videos

Mismatch Repair01:36

Mismatch Repair

Overview
Mismatch Repair01:20

Mismatch Repair

Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
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...
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...
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
Point and Frameshift Mutations01:30

Point and Frameshift Mutations

Point mutations are genetic alterations involving the change of a single nucleotide base pair in DNA. Depending on how the alteration affects protein synthesis, they can lead to various consequences.Point mutations fall into the following types:Silent mutations occur when a nucleotide change does not alter the amino acid sequence due to the redundancy of the genetic code. For instance, changing ACC to ACA still encodes threonine, leaving the protein function unaffected. This occurs because...

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Related Experiment Video

Updated: May 29, 2026

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
06:59

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

Published on: March 31, 2022

Correction of frameshift mutations with tailed duplex DNAs.

Yukiko Morita1, Hiroyuki Tsuchiya, Hideyoshi Harashima

  • 1Faculty of Pharmaceutical Sciences, Hokkaido University, Japan.

Biological & Pharmaceutical Bulletin
|September 2, 2011
PubMed
Summary

Tailed duplex (TD) DNA fragments show potential for correcting frameshift mutations, though efficiencies are currently low. Further optimization is needed for effective gene correction of insertion and deletion mutations.

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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
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11:08

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10:36

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

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Area of Science:

  • Molecular Biology
  • Genetic Engineering
  • Biotechnology

Background:

  • Tailed duplex (TD) DNA fragments are effective in correcting base-substitution mutations.
  • Frameshift mutations, including single-base insertions and deletions, pose challenges for gene correction technologies.

Purpose of the Study:

  • To evaluate the efficacy of TD fragments in correcting single-base insertion (+G) and deletion (-C) frameshift mutations.
  • To compare the gene correction efficiency of TD fragments with single-stranded (ss) DNA fragments for frameshift mutations.

Main Methods:

  • Co-transfection of 5'-TD and 3'-TD DNA fragments with inactivated Hyg-EGFP genes into CHO-K1 cells.
  • Assessment of gene correction efficiency by introducing recovered plasmid DNA into Escherichia coli.
  • Utilizing hygromycin-resistance and enhanced green fluorescent protein (Hyg-EGFP) reporter genes to quantify correction.

Main Results:

  • TD fragments demonstrated relatively low gene correction efficiencies for frameshift mutations compared to base-substitution mutations.
  • Gene correction efficiencies using TD fragments were higher than those achieved with ss DNA fragments.
  • The study identified potential for TD fragments in frameshift mutation correction, albeit with limitations.

Conclusions:

  • TD DNA fragments show promise for correcting frameshift mutations, but require further development for improved efficiency.
  • The findings suggest a basis for future strategies aimed at enhancing TD fragment-mediated gene repair for insertion and deletion mutations.