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Integrated Multimodal Evaluation of Genotoxicity in ZFN-Modified Primary Human Cells

Jaichandran Sivalingam1,2,3, Dimitar Kenanov4, Wai Har Ng5

  • 1Division of Medical Sciences, Laboratory of Applied Human Genetics, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore, Republic of Singapore. jaichandran_sivalingam@bti.a-star.edu.sg.

Methods in Molecular Biology (Clifton, N.J.)
|August 30, 2018
PubMed

Insights

This study introduces a multimodal approach to assess genotoxicity and off-target alterations in gene-edited cells, crucial for ensuring the biosafety of cell therapies and advancing genome editing technologies.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genetics

Background:

  • Clinical trials using retroviral vectors have highlighted the need for rigorous biosafety assessments.
  • Concerns exist regarding genotoxic off-target alterations in gene-corrected cell therapies.

Purpose of the Study:

  • To develop and present an integrated, multimodal approach for investigating off-target alterations.
  • To evaluate the potential genotoxicity of zinc finger nuclease (ZFN)-mediated transgene integration.

Main Methods:

  • Utilized site-specific integration into the AAVS1 locus of primary human cells.
  • Employed a multimodal investigation strategy to detect off-target modifications.
  • Assessed potential genotoxicity associated with ZFN-mediated gene editing.

Main Results:

  • Demonstrated an integrated approach for comprehensive analysis of genome editing outcomes.
  • Provided an evaluation framework for the biosafety of gene-editing technologies.

Conclusions:

  • The developed multimodal approach is essential for stringent assessment of gene-editing safety.
  • This methodology supports the advancement of safer cell-based therapeutic applications.
Keywords:
AAVS1 site-specificCell and gene therapyGenotoxicityOff-targetPrimary human cellsRNA-seq, bioinformaticsTransgene integrationWhole-genome sequencingZinc finger nuclease

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