Genome Editing of Induced Pluripotent Stem Cells Using CRISPR/Cas9 Ribonucleoprotein Complexes to Model Genetic Ocular Diseases

Heran Getachew; Blanca Chinchilla; Rosario Fernandez-Godino

doi:10.1007/7651_2021_409

Genome Editing of Induced Pluripotent Stem Cells Using CRISPR/Cas9 Ribonucleoprotein Complexes to Model Genetic Ocular Diseases

Methods Mol Biol. 2022:2549:321-334. doi: 10.1007/7651_2021_409.

Authors

Heran Getachew¹, Blanca Chinchilla¹, Rosario Fernandez-Godino²

Affiliations

¹ The Ocular Genomics Institute at Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA.
² The Ocular Genomics Institute at Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA. rosario_godino@meei.harvard.edu.

PMID: 34128206
DOI: 10.1007/7651_2021_409

Abstract

Genome editing with the use of CRISPR/Cas9 ribonucleoprotein complexes of induced pluripotent stem cells can be used to model many diseases. The combination of stem cells and gene editing technologies is a valuable tool to study ocular disorders, as many have been identified to be caused by specific genetic mutations. This protocol provides experimentally derived guidelines for genome editing of human induced pluripotent stem cells (iPSCs) using CRISPR/Cas9 ribonucleoprotein complexes to generate iPSCs harboring single nucleotide variants from ocular disorders. Edited iPSC can be further differentiated into retinal cells in order to study disease mechanisms as well as screen potential therapies.

Keywords: CRISPR/Cas9; Genetic ocular disorders; Genome editing; Human iPSC; RNP.

MeSH terms

CRISPR-Cas Systems* / genetics
Cell Differentiation / genetics
Clinical Protocols
Eye Diseases, Hereditary* / genetics
Gene Editing* / methods
Humans
Induced Pluripotent Stem Cells*
Ribonucleoproteins / genetics

Substances

Ribonucleoproteins