University of Pittsburgh

Ira Jacob Fox

Alejandro Soto-Gutierrez

Yang Wang

Yoram Vodovotz

Kan Handa

Kazuki Takeishi

Alexandra Sylvie Collin De l'Hortet

Jorge Guzman Lepe

Aaron W Bell

Taichiro N Nishikawa

Alexandra Sylvie Collin De I'HORTET

Methods are disclosed herein for efficiently generating human induced pluripotent stem cells (iPSC) containing a nucleic acid including a doxycycline promoter operably linked to a nucleic acid encoding Cas9. These methods include transfecting a human somatic cell with a nucleic acid molecule comprising a doxycycline promoter operably linked to a nucleic acid encoding a Cas9, and constitutive promoter operably linked to a tetracycline responsive element and inducing the somatic cell to form an iPSC, thereby producing an iPSC that can undergo CRISPR/Cas9-mediated recombination at a high efficiency. The human iPSC, or a cell differentiated therefrom, is cultured in the presence of doxycycline to induce expression of the Cas9. These cells can then be used to target in any gene of interest by introducing nucleic acids encoding sgRNAs. Induced pluripotent stem cells produced by these methods are also disclosed.

Human induced pluripotent stem cells for high efficiency genetic engineering

The presently disclosed invention is directed to the discovery that hepatocyte nuclear factor 4 alpha (HNF4&#x3b1;; also known as NR2A1), a transcription factor, reverses hepatocyte dysfunction in an animal model of cirrhosis, resulting in improvement in hepatic function, treatment of cirrhosis, and prolonged survival.

Methods for the treatment and prevention of liver disease

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Pittsburgh, PA, United States of America

Ira Jacob Fox