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Related Experiment Video

Updated: May 1, 2026

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Quantifying genome-editing outcomes at endogenous loci with SMRT sequencing.

Ayal Hendel1, Eric J Kildebeck1, Eli J Fine2

  • 1Department of Pediatrics, Stanford University, Stanford, CA 94305, USA.

Cell Reports
|April 2, 2014
PubMed
Summary

Researchers developed a new method using single-molecule real-time (SMRT) DNA sequencing to precisely measure genome editing outcomes. This technique allows simultaneous quantification of various editing events across cell populations for improved gene editing technology evaluation.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Engineered nucleases enable precise genome editing.
  • Measuring editing efficiency and outcomes across cell populations at endogenous loci remains challenging.

Purpose of the Study:

  • To develop a method for simultaneously quantifying diverse genome editing outcomes at any genomic locus.
  • To enable sensitive measurement of editing frequencies in cell populations.

Main Methods:

  • Utilized single-molecule real-time (SMRT) DNA sequencing.
  • Applied the method to various engineered nuclease platforms (TALENs, CRISPR/Cas9, ZFNs) and cell lines.

Main Results:

  • Successfully quantified individual genome editing outcomes at specific genomic sites.
  • Demonstrated applicability across different nuclease platforms and cell types.
  • Identified optimal conditions and strategies for desired genome engineering outcomes.

Conclusions:

  • The developed SMRT sequencing method provides a powerful tool for studying double-strand break repair.
  • Facilitates comprehensive evaluation of gene editing technologies.
  • Enables sensitive quantification of editing outcomes in diverse experimental systems.