Protocol for functional screening of CFTR-targeted genetic therapies in patient-derived organoids using DETECTOR deep-learning-based analysis.

Journal: STAR protocols
PMID:

Abstract

Here, we present a protocol for the rapid functional screening of gene editing and addition strategies in patient-derived organoids using the deep-learning-based tool DETECTOR (detection of targeted editing of cystic fibrosis transmembrane conductance regulator [CFTR] in organoids). We describe steps for wet-lab experiments, image acquisition, and CFTR function analysis by DETECTOR. We also detail procedures for applying pre-trained models and training custom models on new customized datasets. For complete details on the use and execution of this protocol, refer to Bulcaen et al..

Authors

  • Mattijs Bulcaen
    Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, 3000 Leuven, Belgium; Department of Chronic Diseases and Metabolism, KU Leuven, 3000 Leuven, Belgium. Electronic address: bulcaen2@gmail.com.
  • Ronald B Liu
    Department of Biosystems, KU Leuven, 3001 Leuven, Belgium; Institute for Imaging, Data and Communication, University of Edinburgh, Edinburgh EH93JL, UK. Electronic address: bingnan.liu@kuleuven.be.
  • Kasper Gryspeert
    Department of Chronic Diseases and Metabolism, KU Leuven, 3000 Leuven, Belgium. Electronic address: kasper.gryspeert@kuleuven.be.
  • Sam Thierie
    Department of Chronic Diseases and Metabolism, KU Leuven, 3000 Leuven, Belgium.
  • Anabela S Ramalho
    Department of Development and Regeneration, KU Leuven, 3000 Leuven, Belgium.
  • François Vermeulen
    Department of Development and Regeneration, KU Leuven, 3000 Leuven, Belgium; Department of Pediatrics, UZ Leuven, 3000 Leuven, Belgium.
  • Xavier Casadevall I Solvas
    Department of Biosystems, KU Leuven, 3001 Leuven, Belgium.
  • Marianne S Carlon
    Department of Chronic Diseases and Metabolism, KU Leuven, 3000 Leuven, Belgium. Electronic address: marianne.carlon@kuleuven.be.