Optimization of CRISPR/Cas9 Technology to Knock Out Genes of Interest in Aneuploid Cell Lines

Tissue Eng Part C Methods. 2019 Mar;25(3):168-175. doi: 10.1089/ten.TEC.2018.0365.

Abstract

Cell lines represent convenient models to elucidate specific causes of multigenetic and pluricausal diseases, to test breakthrough regenerative technologies. Most commonly used cell lines surpass diploid cells in their accessibility for delivery of large DNA molecules and genome editing, but the main obstacles for obtaining cell models with knockout-targeted protein from aneuploid cells are multiple allele copies and karyotype/phenotype heterogeneity. In the study, we report an original approach to CRISPR-/Cas9-mediated genome modification of aneuploid cell cultures to create functional cell models, achieving highly efficient targeted protein knockout and avoiding "clonal effect" (for the first time to our knowledge).

Keywords: CRISPR/Cas9; cell model; clone effect; genome editing; heterogeneous cell culture; off-targets.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aneuploidy*
  • Animals
  • CRISPR-Cas Systems*
  • Gene Editing*
  • Gene Knockout Techniques / standards*
  • Genes / genetics*
  • HeLa Cells
  • Hep G2 Cells
  • Humans
  • Mice
  • NIH 3T3 Cells