Multiplexed CRISPR/Cas9 targeting of genes implicated in retinal regeneration and degeneration

Arife Unal Eroglu, Timothy S. Mulligan, Liyun Zhang, David T. White, Sumitra Sengupta, Cathy Nie, Noela Y. Lu, Jiang Qian, Lisha Xu, Wuhong Pei, Shawn M. Burgess, Meera T. Saxena, Jeff S. Mumm

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


Thousands of genes have been implicated in retinal regeneration, but only a few have been shown to impact the regenerative capacity of Müller glia-an adult retinal stem cell with untapped therapeutic potential. Similarly, among nearly 300 genetic loci associated with human retinal disease, the majority remain untested in animal models. To address the large-scale nature of these problems, we are applying CRISPR/Cas9-based genome modification strategies in zebrafish to target over 300 genes implicated in retinal regeneration or degeneration. Our intent is to enable large-scale reverse genetic screens by applying a multiplexed gene disruption strategy that markedly increases the efficiency of the screening process. To facilitate large-scale phenotyping, we incorporate an automated reporter quantification-based assay to identify cellular degeneration and regeneration-deficient phenotypes in transgenic fish. Multiplexed gene targeting strategies can address mismatches in scale between "big data" bioinformatics and wet lab experimental capacities, a critical shortfall limiting comprehensive functional analyses of factors implicated in ever-expanding multiomics datasets. This report details the progress we have made to date with a multiplexed CRISPR/Cas9-based gene targeting strategy and discusses how the methodologies applied can further our understanding of the genes that predispose to retinal degenerative disease and which control the regenerative capacity of retinal Müller glia cells.

Original languageEnglish (US)
Article number88
JournalFrontiers in Cell and Developmental Biology
Issue numberAUG
StatePublished - Aug 21 2018


  • CRISPR/Cas9
  • Large scale
  • Multiplex
  • Regeneration
  • Retina
  • Retinal degeneration
  • Rhodopsin
  • Zebrafish

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology


Dive into the research topics of 'Multiplexed CRISPR/Cas9 targeting of genes implicated in retinal regeneration and degeneration'. Together they form a unique fingerprint.

Cite this