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

Avellino Lab USA

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

2 Citations (Scopus)

162 Downloads (Pure)

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

UN SDGs

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

SDG 3 Good Health and Well-being

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)

Access to Document

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

e.Proofing _ SpringerAuthor Accepted Manuscript, 724 KB

Cite this

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title = "Gene Editing for Corneal Stromal Regeneration",

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.",

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T1 - Gene Editing for Corneal Stromal Regeneration

AU - Moore, Tara C. B.

AU - Chao-Shern, Connie

AU - DeDionisio, Larry

AU - Christie, Kathleen A.

AU - Nesbit, M. Andrew

PY - 2020/6/16

Y1 - 2020/6/16

N2 - 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.

AB - 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.

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)

UR - https://www.scopus.com/pages/publications/85086621907

UR - https://www.springer.com/gp/book/9781071605981?utm_campaign=bookpage_about_buyonpublisherssite&utm_medium=referral&utm_source=springerlink

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

M3 - Chapter

C2 - 32542601

SN - 978-1-0716-0598-1

VL - 2145

T3 - Methods in Molecular Biology

SP - 59

EP - 75

BT - Corneal Regeneration

A2 - Ahearne, Mark

PB - Springer Nature

CY - New York, NY

ER -

Gene Editing for Corneal Stromal Regeneration

Abstract

Publication series

UN SDGs

Keywords

Access to Document

Other files and links

Fingerprint

Development of partner diagnostics and therapeutics for genetic eye disease

Cite this

Gene Editing for Corneal Stromal Regeneration

Abstract

Publication series

UN SDGs

Keywords

Access to Document

Other files and links

Fingerprint

Student theses

Development of partner diagnostics and therapeutics for genetic eye disease

Cite this