Please use this identifier to cite or link to this item: https://doi.org/10.1155/2016/3598542
Title: Targeted Inhibition of the MIR-199a/214 Cluster by CRISPR Interference Augments the Tumor Tropism of Human Induced Pluripotent Stem Cell-Derived Neural Stem Cells under Hypoxic Condition
Authors: Luo, Y
Xu, X
An, X
Sun, X
Wang, S 
Zhu, D
Keywords: hypoxia inducible factor 1alpha
microRNA
microRNA 199a
microRNA 199a 3p
microRNA 199a 5p
microRNA 214
mitogen activated protein kinase 1
scatter factor receptor
unclassified drug
animal cell
animal experiment
animal model
Article
cell hypoxia
cell migration
clustered regularly interspaced short palindromic repeat
controlled study
female
gene cluster
gene expression
human
human cell
in vitro study
in vivo study
induced pluripotent stem cell
mouse
neural stem cell
nonhuman
promoter region
tropism
upregulation
Issue Date: 2016
Citation: Luo, Y, Xu, X, An, X, Sun, X, Wang, S, Zhu, D (2016). Targeted Inhibition of the MIR-199a/214 Cluster by CRISPR Interference Augments the Tumor Tropism of Human Induced Pluripotent Stem Cell-Derived Neural Stem Cells under Hypoxic Condition. Stem Cells International 2016 : 3598542. ScholarBank@NUS Repository. https://doi.org/10.1155/2016/3598542
Rights: Attribution 4.0 International
Abstract: The human induced pluripotent stem cell (hiPSC) provides a breakthrough approach that helps overcoming ethical and allergenic challenges posed in application of neural stem cells (NSCs) in targeted cancer gene therapy. However, the tumor-tropic capacity of hiPSC-derived NSCs (hiPS-NSCs) still has much room to improve. Here we attempted to promote the tumor tropism of hiPS-NSCs by manipulating the activity of endogenous miR-199a/214 cluster that is involved in regulation of hypoxia-stimulated cell migration. We first developed a baculovirus-delivered CRISPR interference (CRISPRi) system that sterically blocked the E-box element in the promoter of the miR-199a/214 cluster with an RNA-guided catalytically dead Cas9 (dCas9). We then applied this CRISPRi system to hiPS-NSCs and successfully suppressed the expression of miR-199a-5p, miR-199a-3p, and miR-214 in the microRNA gene cluster. Meanwhile, the expression levels of their targets related to regulation of hypoxia-stimulated cell migration, such as HIF1A, MET, and MAPK1, were upregulated. Further migration assays demonstrated that the targeted inhibition of the miR-199a/214 cluster significantly enhanced the tumor tropism of hiPS-NSCs both in vitro and in vivo. These findings suggest a novel application of CRISPRi in NSC-based tumor-targeted gene therapy. © 2016 Yumei Luo et al.
Source Title: Stem Cells International
URI: https://scholarbank.nus.edu.sg/handle/10635/179957
ISSN: 16879678
DOI: 10.1155/2016/3598542
Rights: Attribution 4.0 International
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