SINGLE LENTIVIRAL VECTOR SYSTEM FOR INDUCED PLURIPOTENT (IPS) STEM CELLS DERIVATION

摘要

The present invention is based on the discovery that a single lentiviral vector expressing multiple individual transcription factor proteins from a single multi-cistronic mRNA can reprogram a fibroblast cell to a stem cell-like cell. These reprogrammed induced pluripotent stem (iPS) cells are pluripotent. Additions of the Cre-LoxP sequences into the single lentiviral vector facilitate excision of the vector after reprogramming in achieved. Addition of a maker gene into the single lentiviral vector facilitates detection of the presence of the vector in an iPS. The invention provides compositions and methods of producing iPS cells using a single multi-cistronic lentiviral vector.

主权项

1. A lentiviral vector comprising a nucleic acid sequence operatively linked to a promoter, wherein the nucleic acid sequences comprises:
a. a ‘self-cleaving’ 2A peptide; b. an internal ribosome entry site (IRES); and c. a first gene, a second gene, and third gene selected from the group of genes consisting of Oct4, Klf4 and Sox2; wherein the first gene, second gene and third gene can be in any order and wherein the ‘self-cleaving’ 2A peptide is positioned between the first gene and second gene, and wherein the IRES is positioned between the second and third genes, and wherein the nucleic acid sequence encoding the first gene, second gene and third gene, the ‘self-cleaving’ 2A peptide and the IRES are transcribed from the promoter as a multi-cistronic RNA.