Pubblicazione scientifica

Multiplexed CRISPR/Cas9 Targeting of Genes Implicated in Retinal Regeneration and Degeneration

FILONE TECNOLOGICO	TEMA
produzione e raccolta

RIASSUNTO

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.

DATA

Data di pubblicazione: 21/08/2018

AUTORI

UNAL EROGLU ARIFE
MULLIGAN TIMOTHY S
ZHANG LIYUN
WHITE DAVID T
SENGUPTA SUMITRA
NIE CATHY
LU NOELA Y
QIAN JIANG
XU LISHA
PEI WUHONG
BURGESS SHAWN M
SAXENA MEERA T
MUMM JEFF S

ENTE DI AFFERENZA

DEPT OPHTHALMOLOGY WILMER EYE INST JOHNS HOPKINS UNIV SCHOOL OF MEDICINE BALTIMORE MD US
TRANSLATIONAL & FUNCTIONAL GENOMICS BRANCH NATL HUMAN GENOME RSCH INST BETHESDA MD US

RIVISTA

Frontiers in Cell and Developmental Biology, 6, 88

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