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 proceedingChapter

1 Citation (Scopus)
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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 languageEnglish
Title of host publicationCorneal Regeneration
Subtitle of host publicationMethods in Molecular Biology
EditorsMark Ahearne
Place of PublicationNew York, NY
PublisherSpringer Nature
Number of pages17
ISBN (Electronic)978-1-0716-0599-8
ISBN (Print)978-1-0716-0598-1
Publication statusPublished online - 16 Jun 2020

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • 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)


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