Integrating CRISPR Engineering and hiPSC-Derived 2D Disease Modeling Systems

Abstract

Human induced pluripotent stem cells (hiPSCs) have revolutionized research on human diseases, particularly neurodegenerative and psychiatric disorders, making it possible to study mechanisms of disease risk and initiation in otherwise inaccessible patient-specific cells. Today, the integration of CRISPR engineering approaches with hiPSC-based models permits precise isogenic comparisons of human neurons and glia. This review is intended as a guideline for neuroscientists and clinicians interested in translating their research to hiPSC-based studies. It offers state-of-the-art approaches to tackling the challenges that are unique to human in vitro disease models, particularly interdonor and intradonor variability, and limitations in neuronal maturity and circuit complexity. Finally, we provide a detailed overview of the immense possibilities the field has to offer, highlighting efficient neural differentiation and induction strategies for the major brain cell types and providing perspective into integrating CRISPR-based methods into study design. The combination of hiPSC-based disease modeling, CRISPR technology, and high-throughput approaches promises to advance our scientific knowledge and accelerate progress in drug discovery.Dual Perspectives Companion Paper: Studying Human Neurodevelopment and Diseases Using 3D Brain Organoids, by Ai Tian, Julien Muffat, and Yun Li.

Keywords: CRISPR; Human induced pluripotent stem cells; differentiation; disease modeling; induction; psychiatric genetics.

Figure 1.

Approaching hiPSC disease modeling. The general human population exhibits high genetic variability, which can be overcome by stratification of the cohort or by CRISPR editing approaches. Generated iPSC lines can be differentiated or induced into several neural cell types, comprising neurons and glia, which can be modularly combined into cocultures for complex genetic, phenotypic, or functional analysis. Components of schematic were adapted from Servier Medical Art (https://smart.servier.com/#) and BioRender (created with www.BioRender.com).