Stem Cell Reports
Volume 9, Issue 2, 8 August 2017, Pages 615-628
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Article
Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes

https://doi.org/10.1016/j.stemcr.2017.06.012Get rights and content
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Highlights

  • The efficacy of CRISPR-mediated transcript modulation varies between genes

  • gRNAs should be re-validated for each individual, cell type, and dCas9-effector

Summary

Modulation of transcription, either synthetic activation or repression, via dCas9-fusion proteins is a relatively new methodology with the potential to facilitate high-throughput up- or downregulation studies of gene function. Genetic studies of neurodevelopmental disorders have identified a growing list of risk variants, including both common single-nucleotide variants and rare copy-number variations, many of which are associated with genes having limited functional annotations. By applying a CRISPR-mediated gene-activation/repression platform to populations of human-induced pluripotent stem cell-derived neural progenitor cells, neurons, and astrocytes, we demonstrate that it is possible to manipulate endogenous expression levels of candidate neuropsychiatric risk genes across these three cell types. Although proof-of-concept studies using catalytically inactive Cas9-fusion proteins to modulate transcription have been reported, here we present a detailed survey of the reproducibility of gRNA positional effects across a variety of neurodevelopmental disorder-relevant risk genes, donors, neural cell types, and dCas9 effectors.

Keywords

CRISPR
human-induced pluripotent stem cell
neural progenitor cell
transcriptional modulation
dCas9-VP64
dCas9-VPR
dCas9-KRAB

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Co-first author