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Blue Light‐Operated CRISPR/Cas13b‐Mediated mRNA Knockdown (Lockdown)
Author(s) -
Blomeier Tim,
Fischbach Patrick,
Koch LeonieAlexa,
Andres Jennifer,
Miñambres Miguel,
Beyer Hannes Michael,
Zurbriggen Matias Daniel
Publication year - 2021
Publication title -
advanced biology
Language(s) - English
Resource type - Journals
ISSN - 2701-0198
DOI - 10.1002/adbi.202000307
Subject(s) - gene knockdown , optogenetics , rna interference , crispr , downregulation and upregulation , microbiology and biotechnology , messenger rna , biology , crispr interference , gene expression , rna , regulation of gene expression , gene , cas9 , genetics , neuroscience
The introduction of optogenetics into cell biology has furnished systems to control gene expression at the transcriptional and protein stability level, with a high degree of spatial, temporal, and dynamic light‐regulation capabilities. Strategies to downregulate RNA currently rely on RNA interference and CRISPR/Cas‐related methods. However, these approaches lack the key characteristics and advantages provided by optical control. “Lockdown” introduces optical control of RNA levels utilizing a blue light‐dependent switch to induce expression of CRISPR/Cas13b, which mediates sequence‐specific mRNA knockdown. Combining Lockdown with optogenetic tools to repress gene‐expression and induce protein destabilization with blue light yields efficient triple‐controlled downregulation of target proteins. Implementing Lockdown to degrade endogenous mRNA levels of the cyclin‐dependent kinase 1 (hCdk1) leads to blue light‐induced G2/M cell cycle arrest and inhibition of cell growth in mammalian cells.