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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 :
Yumei Luo
Xuehu Xu
Xiuli An
Xiaofang Sun
Shu Wang
Detu Zhu
Source :
Stem Cells International, Vol 2016 (2016)
Publication Year :
2016
Publisher :
Hindawi Limited, 2016.

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.

Subjects

Subjects :
Internal medicine
RC31-1245

Details

Language :
English
ISSN :
1687966X and 16879678
Volume :
2016
Database :
Directory of Open Access Journals
Journal :
Stem Cells International
Publication Type :
Academic Journal
Accession number :
edsdoj.8422d3090a904c179f6c73c3d148551c
Document Type :
article
Full Text :
https://doi.org/10.1155/2016/3598542