Details

Autor(en) / Beteiligte

• Gallego-Perez, Daniel, PhD
• Otero, Jose J., MD, PhD
• Czeisler, Catherine, PhD
• Ma, Junyu, PhD
• Ortiz, Cristina, BSc
• Gygli, Patrick, PhD
• Catacutan, Fay Patsy, BSc
• Gokozan, Hamza Numan, MD
• Cowgill, Aaron
• Sherwood, Thomas, PhD
• Ghatak, Subhadip, PhD
• Malkoc, Veysi, PhD
• Zhao, Xi, PhD
• Liao, Wei-Ching, PhD
• Gnyawali, Surya, PhD
• Wang, Xinmei, PhD
• Adler, Andrew F., PhD
• Leong, Kam, PhD
• Wulff, Brian, PhD
• Wilgus, Traci A., PhD
• Askwith, Candice, PhD
• Khanna, Savita, PhD
• Rink, Cameron, PhD
• Sen, Chandan K., PhD
• Lee, L. James, PhD

Titel

Deterministic transfection drives efficient nonviral reprogramming and uncovers reprogramming barriers

Ist Teil von

• Nanomedicine, 2016-02, Vol.12 (2), p.399-409

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• Abstract Safety concerns and/or the stochastic nature of current transduction approaches have hampered nuclear reprogramming's clinical translation. We report a novel non-viral nanotechnology-based platform permitting deterministic large-scale transfection with single-cell resolution. The superior capabilities of our technology are demonstrated by modification of the well-established direct neuronal reprogramming paradigm using overexpression of the transcription factors Brn2 , Ascl1 , and Myt1l (BAM). Reprogramming efficiencies were comparable to viral methodologies (up to ~ 9-12%) without the constraints of capsid size and with the ability to control plasmid dosage, in addition to showing superior performance relative to existing non-viral methods. Furthermore, increased neuronal complexity could be tailored by varying BAM ratio and by including additional proneural genes to the BAM cocktail. Furthermore, high-throughput NEP allowed easy interrogation of the reprogramming process. We discovered that BAM-mediated reprogramming is regulated by AsclI dosage, the S-phase cyclin CCNA2 , and that some induced neurons passed through a nestin-positive cell stage.