Integration-free human induced pluripotent stem cells from...

Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...

Reexamination Certificate

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C435S320100, C435S325000

Reexamination Certificate

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08048675

ABSTRACT:
Provided herein are methods for generating human induced pluripotent stem cells free from genomic integration of exogenous transgenes by transfecting into nucleated blood cells one or more DNA expression vectors (e.g., plasmid vectors) that do not contain a mammalian origin of replication, and encode and permit expression of one or more reprogramming factors (e.g., Oct4, Sox2, Klf4, and c-Myc). Also provided herein are the integration-free human induced pluripotent stem cells obtained by the methods described herein.

REFERENCES:
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patent: 2010/0003757 (2010-01-01), Mack et al.
patent: WO 2009/115295 (2009-09-01), None
patent: WO 2009/133971 (2009-11-01), None
patent: WO 2009/157201 (2009-12-01), None
patent: WO 2010/012077 (2010-02-01), None
Yusa et al (Nature Method, 2009. vol. 6, No. 5, pp. 363-369).
Jia, et al. A nonviral minicircle vector for deriving human iPS cells. Nat Methods. Mar. 2010;7(3):197-9.
Kaji, et al. Virus-free induction of pluripotency and subsequent excision of reprogramming factors. Nature. Apr. 9, 2009;458(7239):771-5.
Okita, et al. Generation of mouse induced pluripotent stem cells without viral vectors. Science. Nov. 7, 2008;322(5903):949-53.
Woltjen, et al. piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells. Nature. Apr. 9, 2009;458(7239):766-70.
Yu, et al. Human induced pluripotent stem cells free of vector and transgene sequences. Science. May 8, 2009;324(5928):797-801.

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