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Genetic spell-checking: gene editing using single-stranded DNA oligonucleotides.

Natalia Rivera-Torres1, Eric B Kmiec1

  • 1Gene Editing Institute, Center for Translational Cancer Research, Helen F. Graham Cancer Center & Research Institute, Christiana Care Health System, Newark, DE, USA.

Plant Biotechnology Journal
|September 25, 2015
PubMed
Summary
This summary is machine-generated.

Single-stranded oligonucleotides (ssODNs) facilitate gene editing by leveraging cellular repair pathways. Understanding DNA damage response and replication fork stalling can enhance ssODN-mediated gene modification efficiency in plants.

Keywords:
gene editingmolecular geneticssingle-stranded oligonucleotides

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Area of Science:

  • Molecular Biology
  • Plant Science
  • Genetics

Background:

  • Gene editing utilizes single-stranded oligonucleotides (ssODNs) for precise genetic alterations.
  • ssODNs, composed of DNA or mixed RNA/DNA bases, exploit cellular metabolic functions for gene modification.
  • Elucidating gene editing mechanisms is crucial for developing effective plant gene editing strategies.

Purpose of the Study:

  • To discuss critical reaction parameters influencing gene editing effectiveness.
  • To present functional interrelationships between DNA damage, DNA response pathways, and replication fork stalling.
  • To identify potential targets for increasing gene editing frequency by ssODNs in plants.

Main Methods:

  • Review of current understanding of gene editing mechanisms.
  • Analysis of the role of DNA damage and repair pathways.
  • Examination of replication fork dynamics in relation to gene editing.

Main Results:

  • Double-strand DNA breaks and DNA damage response activation are key determinants of gene editing frequency.
  • Cellular regulators responding to DNA damage influence the success of nucleotide exchange reactions.
  • Replication fork stalling, induced by DNA damage response, increases chromatin accessibility for ssODNs.

Conclusions:

  • Gene editing efficiency is modulated by the interplay of DNA damage, repair pathways, and replication fork progression.
  • Targeting these pathways offers a strategy to enhance ssODN-mediated gene editing in plants and plant cells.