Gene Editing for Corneal Stromal Regeneration

Tara C. B. Moore; Connie Chao-Shern; Larry DeDionisio; Kathleen A. Christie; M. Andrew Nesbit

doi:10.1007/978-1-0716-0599-8_6

Gene Editing for Corneal Stromal Regeneration

Tara C. B. Moore, Connie Chao-Shern, Larry DeDionisio, Kathleen A. Christie, M. Andrew Nesbit

Research output: Chapter in Book/Report/Conference proceeding › Chapter

1 Citation (Scopus)

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Abstract

CRISPR/Cas9 gene editing holds the promise of sequence-specific alteration of the genome to achieve therapeutic benefit in the treated tissue. Cas9 is an RNA-guided nuclease in which the sequence of the RNA can be altered to match the desired target. However, care must be taken in target choice and RNA guide design to ensure both maximum on-target and minimum off-target activity. The cornea is an ideal tissue for gene therapy due to its small surface area, accessibility, immune privilege, avascularity, and ease of visualization. Herein, we describe the design, testing, and delivery of Cas9 and guide RNAs to target genes expressed in the cornea.

Original language	English
Title of host publication	Corneal Regeneration
Subtitle of host publication	Methods in Molecular Biology
Editors	Mark Ahearne
Place of Publication	New York, NY
Publisher	Springer Nature
Chapter	6
Pages	59-75
Number of pages	17
Volume	2145
ISBN (Electronic)	978-1-0716-0599-8
ISBN (Print)	978-1-0716-0598-1
DOIs	https://doi.org/10.1007/978-1-0716-0599-8_6
Publication status	Published online - 16 Jun 2020

Publication series

Name	Methods in Molecular Biology
Volume	2145
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

Gene editing
sgRNA expression construct
Dual luciferase assay
In vitro digest
Lymphoblastoid cell line
Nucleofection
Polymerase chain reaction (PCR)
Corneal endothelial and epithelial cell culture
In vivo imaging
Intracameral injection
Adeno-associated virus (AAV)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/978-1-0716-0599-8_6

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Cite this

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AU - DeDionisio, Larry

AU - Christie, Kathleen A.

AU - Nesbit, M. Andrew

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KW - Gene editing

KW - sgRNA expression construct

KW - Dual luciferase assay

KW - In vitro digest

KW - Lymphoblastoid cell line

KW - Nucleofection

KW - Polymerase chain reaction (PCR)

KW - Corneal endothelial and epithelial cell culture

KW - In vivo imaging

KW - Intracameral injection

KW - Adeno-associated virus (AAV)

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ER -

Gene Editing for Corneal Stromal Regeneration

Abstract

Publication series

Keywords

UN SDGs

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