CRISPR/Cas9 Gene Editing In Vitro and in Retinal Cells In Vivo

Methods Mol Biol. 2019:1834:59-74. doi: 10.1007/978-1-4939-8669-9_4.


CRISPR/Cas9 is an efficient tool to knock down specific genes in various organisms. In this chapter, we describe how to assess knockdown of human rhodopsin (RHO) gene carrying the P23H mutation in vitro, in engineered HeLa cells, and in vivo, in P23H RHO transgenic mice. To this aim, we report two molecular assays: site-specific PCR on P23H RHO cells treated with CRISPR/Cas9 and Western blotting analysis on retinal cells prepared from P23H RHO transgenic mice electroporated with CRISPR/Cas9 and GFP plasmids.

Keywords: CRISPR/Cas9 plasmid; Knockdown; Retinal cells; Rhodopsin gene; Site-specific PCR; Western blotting.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Cloning, Molecular
  • Computational Biology / methods
  • Fluorescent Antibody Technique
  • Gene Editing*
  • Gene Expression
  • Gene Knockdown Techniques
  • Gene Targeting
  • HeLa Cells
  • Humans
  • Mice
  • Polymerase Chain Reaction
  • RNA, Guide, Kinetoplastida
  • Retina / cytology*
  • Retina / metabolism*
  • Rhodopsin / genetics


  • RNA, Guide
  • Rhodopsin