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

STAR Protoc. 2025 Mar 21;6(1):103593. doi: 10.1016/j.xpro.2024.103593. Epub 2025 Jan 31.

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.1.

Keywords: CRISPR; computer sciences; organoids.

MeSH terms

  • Cystic Fibrosis / genetics
  • Cystic Fibrosis / therapy
  • Cystic Fibrosis Transmembrane Conductance Regulator* / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator* / metabolism
  • Deep Learning*
  • Gene Editing* / methods
  • Genetic Therapy* / methods
  • Humans
  • Organoids* / metabolism

Substances

  • Cystic Fibrosis Transmembrane Conductance Regulator
  • CFTR protein, human