title: Generation of induced pluripotent stem cells by using a mammalian artificial chromosome expression system.
abstract: Direct reprogramming of mouse fibroblasts into induced pluripotent stem cells (iPS) was achieved recently by overexpression of four transcription factors encoded by retroviral vectors. Most of the virus vectors, however, may cause insertional mutagenesis in the host genome and may also induce tumor formation. Therefore, it is very important to discover novel and safer, non-viral reprogramming methods. Here we describe the reprogramming of somatic cells into iPS cells by a novel protein-based technique. Engineered Oct4, Sox2 and Klf4 transcription factors carrying an N-terminal Flag-tag and a C-terminal polyarginine tail were synthesized by a recently described mammalian artificial chromosome expression system (ACEs). This system is suitable for the high-level production of recombinant proteins in mammalian tissue culture cells. Recombinant proteins produced in this system contain all the post-translational modifications essential for the stability and the authentic function of the proteins. The engineered Oct4, Sox2 and Klf4 proteins efficiently induced the reprogramming of mouse embryonic fibroblasts by means of protein transduction. This novel method allows for the generation of iPS cells, which may be suitable for therapeutic applications in the future.
authors.affiliation: Hungarian Academy of Sciences Institute of Genetics, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: Anna
authors.initials: A
authors.lastname: Tóth
authors.affiliation: Hungarian Academy of Sciences Institute of Genetics, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: Katalin
authors.initials: K
authors.lastname: Fodor
authors.affiliation: Hungarian Academy of Sciences Institute of Genetics, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: P
authors.initials: P
authors.lastname: Blazsó
authors.affiliation: Hungarian Academy of Sciences Institute of Genetics, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: I
authors.initials: I
authors.lastname: Cserpán
authors.affiliation: Hungarian Academy of Sciences Institute of Genetics, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: Tünde
authors.initials: T
authors.lastname: Praznovszky
authors.affiliation: Hungarian Academy of Sciences Institute of Genetics, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: V
authors.initials: V
authors.lastname: Tubak
authors.affiliation: Hungarian Academy of Sciences Institute of Biochemistry, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: A
authors.initials: A
authors.lastname: Udvardy
authors.affiliation: Hungarian Academy of Sciences Institute of Genetics, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: Gy
authors.initials: G
authors.lastname: Hadlaczky
authors.affiliation: Hungarian Academy of Sciences Institute of Genetics, Biological Research Centre Temesvári krt. 62 H-6726 Szeged Hungary.
authors.forename: R L
authors.initials: RL
authors.lastname: Katona
mesh_terms: D000818:Animals; D016466:CHO Cells; D065150:Cellular Reprogramming; D022221:Chromosomes, Artificial, Mammalian; D018920:Coculture Techniques; D006224:Cricetinae; D003412:Cricetulus; D005347:Fibroblasts / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
chemical_list: C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors
keywords: Artificial Chromosome Expression system; cell and gene therapy; iPS cell; reprogramming; reprogramming proteins