scholarly journals An Integrated Miniature Bioprocessing for Personalized Human Induced Pluripotent Stem Cell Expansion and Differentiation into Neural Stem Cells

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Haishuang Lin ◽  
Qiang Li ◽  
Yuguo Lei
2019 ◽  
Vol 12 ◽  
pp. 88-93 ◽  
Author(s):  
Vytautas Galvanauskas ◽  
Rimvydas Simutis ◽  
Suman Chandra Nath ◽  
Masahiro Kino-oka

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Yumei Luo ◽  
Xuehu Xu ◽  
Xiuli An ◽  
Xiaofang Sun ◽  
Shu Wang ◽  
...  

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.


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