Protocols for Performing CRISPR Gene Editing in Anterior Chamber Eye Cells
- Authors: Sam Yacoub1, Charles E. Amankwa2, Gulab Zode3
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View Affiliations Hide Affiliations1 Department of Pharmacology and Neuroscience and The North Texas Eye Research Institute, University of North Texas Health Science Center, Fort Worth, TX 76107, USA 2 Department of Pharmacology and Neuroscience and The North Texas Eye Research Institute, University of North Texas Health Science Center, Fort Worth, TX 76107, USA 3 Department of Pharmacology and Neuroscience and The North Texas Eye Research Institute, University of North Texas Health Science Center, Fort Worth, TX 76107, USA
- Source: Research Protocols for Ophthalmic Disease Mechanisms and Therapeutics: Glaucoma - Ocular Hypertension , pp 550-564
- Publication Date: August 2025
- Language: English
Protocols for Performing CRISPR Gene Editing in Anterior Chamber Eye Cells, Page 1 of 1
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The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology has revolutionized genetics and opened up new possibilities for research and therapy. The CRISPR system is composed of single guide RNA (sgRNA) and the endonuclease (Cas9), which can be combined into a ribonucleoprotein complex (RNP). RNP can be used to target and cut specific DNA regions. The introduction of foreign DNA at the cut's site allows for precise genome editing, such as insertion, deletion, or substitution of specific genes. This powerful technology has been widely adopted in many fields, including basic research, drug discovery, and biotechnology. Its ability to efficiently edit genes has made it an essential tool for studying gene function, developing new therapies for genetic diseases, and improving crops and livestock. This chapter will discuss the utility of CRISPR/Cas9 and sgRNA using trabecular meshwork Cells (GTM3 cells), highlighting the limitations and potential alternative methods. We will also describe the T7 endonuclease I (T7EI) assay, a protocol used to confirm gene editing.
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