CRISPR engineering in organoids for gene repair and disease modelling

Abstract

Organoids bridge the gap between 2D cell lines and in vivo studies. With their 3D organization and cellular heterogeneity, adult stem cell-derived organoids closely resemble their tissue of origin. The development of CRISPR-mediated genome engineering and the recent additions of base and prime editing to the CRISPR toolbox have greatly simplified the generation of exact, isogenic models for Mendelian diseases. Here, we review recent developments in CRISPR-mediated genome engineering and its application in human adult-stem-cell-derived organoids in the construction of isogenic disease models. These models allow accurate qualification of the impact of allelic disease variants observed in patients. Furthermore, we discuss the use of organoids as models for safety and efficacy of CRISPR for gene repair. Although transplantation of repaired tissue remains challenging, benchmarking CRISPR tools in organoids can bring genome engineering one step closer to patients. Adult stem cell-derived organoids closely resemble their tissue of origin. This Review discusses recent developments in CRISPR-mediated genome engineering and its application using adult-stem-cell-derived organoids in the construction of isogenic disease models and for clinical gene repair.