Efficient genetic editing of human intestinal organoids using ribonucleoprotein-based CRISPR.

ABSTRACT: Organoids, combined with genetic editing strategies, have the potential to offer rapid and efficient investigation of gene function in many models of human disease. However, to date, the editing efficiency of organoids with the use of non-viral electroporation methods has only been up to 30%, with implications for the subsequent need for selection, including turnaround time and exhaustion or adaptation of the organoid population. Here, we describe an efficient method for intestinal organoid editing using a ribonucleoprotein-based CRISPR approach. Editing efficiencies of up to 98% in target genes were robustly achieved across different gut anatomical locations and developmental timepoints from multiple patient samples with no observed off-target editing. The method allowed us to study the e

SUBMITTER: Skoufou-Papoutsaki N 

PROVIDER: S-EPMC10565108 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature