Understanding neural development and diseases using CRISPR screens in human pluripotent stem cell-derived cultures

Mai Ahmed; Julien Muffat; Yun Li

doi:10.3389/fcell.2023.1158373

Understanding neural development and diseases using CRISPR screens in human pluripotent stem cell-derived cultures

Front Cell Dev Biol. 2023 Apr 10:11:1158373. doi: 10.3389/fcell.2023.1158373. eCollection 2023.

Authors

Mai Ahmed¹, Julien Muffat^{2

3}, Yun Li^{1

3}

Affiliations

¹ Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada.
² Program in Neurosciences and Mental Health, The Hospital for Sick Children, Toronto, ON, Canada.
³ Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.

Abstract

The brain is arguably the most complex part of the human body in form and function. Much remains unclear about the molecular mechanisms that regulate its normal and pathological physiology. This lack of knowledge largely stems from the inaccessible nature of the human brain, and the limitation of animal models. As a result, brain disorders are difficult to understand and even more difficult to treat. Recent advances in generating human pluripotent stem cells (hPSCs)-derived 2-dimensional (2D) and 3-dimensional (3D) neural cultures have provided an accessible system to model the human brain. Breakthroughs in gene editing technologies such as CRISPR/Cas9 further elevate the hPSCs into a genetically tractable experimental system. Powerful genetic screens, previously reserved for model organisms and transformed cell lines, can now be performed in human neural cells. Combined with the rapidly expanding single-cell genomics toolkit, these technological advances culminate to create an unprecedented opportunity to study the human brain using functional genomics. This review will summarize the current progress of applying CRISPR-based genetic screens in hPSCs-derived 2D neural cultures and 3D brain organoids. We will also evaluate the key technologies involved and discuss their related experimental considerations and future applications.

Keywords: CRISPR; genetic screens; human pluripotent stem cell (hPSC); neurodevelopment; neurological disease; organoids.

Publication types

Review

Grants and funding

This work was supported by the Canada Research Chairs Program, Brain Canada, Medicine by Design, Brain and Behaviour Research Foundation, the Simons Foundation, and the Natural Science and Engineering Research Council.