Journal
STEM CELLS
Volume 29, Issue 11, Pages 1684-1695Publisher
WILEY
DOI: 10.1002/stem.726
Keywords
microRNA; Induced pluripotent stem cells; Genetic reprogramming; Let7a; Lin28; Parkinson's disease; Rett syndrome
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Funding
- Ministry of Health
- EU (ERANET-Neuron)
- Cariplo Foundation
- Italian Institute of Technology (IIT)
- Telethon
- EU [FP5 LSHB-CT-2004-005242 CONSERT, FP7 GA 222878 PERSIST]
- EU (ERC) [249845 TARGETINGGENETHERAPY]
- Fondazione CARIPLO (NOBEL)
- Italian Ministry of Health
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Induced pluripotent stem cell (iPSC) technology has provided researchers with a unique tool to derive disease-specific stem cells for the study and possible treatment of degenerative disorders with autologous cells. The low efficiency and heterogeneous nature of reprogramming is a major impediment to the generation of personalized iPSC lines. Here, we report the generation of a lentiviral system based on a microRNA-regulated transgene that enables for the efficient selection of mouse and human pluripotent cells. This system relies on the differential expression pattern of the mature form of microRNA let7a in pluripotent versus committed or differentiated cells. We generated microRNA responsive green fluorescent protein and Neo reporters for specific labeling and active selection of the pluripotent cells in any culture condition. We used this system to establish Rett syndrome and Parkinson's disease human iPSCs. The presented selection procedure represents a straightforward and powerful tool for facilitating the derivation of patient-specific iPSCs. STEM CELLS 2011;29:1684-1695
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