The disclosure relates to a method of reprogramming one or more somatic cells, e.g., partially differentiated or fully/terminally differentiated somatic cells, to a less differentiated state, e.g., a pluripotent or multipotent state. In further embodiments the invention also relates to reprogrammed somatic cells produced by methods of the invention, to chimeric animals comprising reprogrammed somatic cells of the invention, to uses of said cells, and to methods for identifying agents useful for reprogramming somatic cells.

An episomal plasmid comprising a gene of interest (GOI) and an autonomously replicating sequence (ARS) which is not operably linked to the GOI, which ARS comprises or consists of a nucleotide sequence identified as any of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:6-11, or a functionally active variant of any of the foregoing which is characterized by at least 60% sequence identity thereto.

The present invention provides HIV-derived lentivectors which are safe, highly efficient, and very potent for expressing transgenes for human gene therapy, especially, in human hematopoietic progenitor cells as well as in all other blood cell derivatives. The lentiviral vectors comprise a self-inactivating configuration for biosafety and promoters such as the EF1? promoter as one example. Additional promoters are also described. The vectors can also comprise additional transcription enhancing elements such as the wood chuck hepatitis virus post-transcriptional regulatory element. These vectors therefore provide useful tools for genetic treatments such as inherited and acquired lympho-hematological disorders, gene-therapies for cancers especially the hematological cancers, as well as for the study of hematopoiesis via lentivector-mediated modification of human HSCs.

Methods and compositions relating to the production of induced pluripotent stem cells (iPS cells) are disclosed. For example, induced pluripotent stem cells may be generated from CD34.sup.+ hematopoietic cells, such as human CD34.sup.+ blood progenitor cells, or T cells. Various iPS cell lines are also provided. In certain embodiments, the invention provides novel induced pluripotent stem cells with a genome comprising genetic rearrangement of T cell receptors.

The present invention relates to a non-viral iPSCs induction method as well as the compositions, kits and iPSCs obtained therefrom. More specifically, the induction method comprises the following steps: 1) Constructing a recombinant plasmid by introducing the DNA sequences expressing the reprogramming factors POU5F1, SOX2, GLIS1, KLF4, MYCL and hsa-miR-302s into an episomal vector; 2) Obtaining iPSCs by introducing the recombinant plasmids obtained in step 1) into human somatic cells, and reprogramming induction culture of the cells. The method reduces the risk of clinical applications of iPSCs by using a combination of highly-safe reprogramming factors without the introduction of high-risk reprogramming factors such as c-MYC, SV40-LT and TP53 inhibitors; The method is highly applicable.

The present invention relates to a non-viral iPSCs induction method as well as the compositions, kits and iPSCs obtained therefrom. More specifically, the induction method comprises the following steps: 1) Constructing a recombinant plasmid by introducing the DNA sequences expressing the reprogramming factors POU5F1, SOX2, GLIS1, KLF4, MYCL and hsa-miR-302s into an episomal vector; 2) Obtaining iPSCs by introducing the recombinant plasmids obtained in step 1) into human somatic cells, and reprogramming induction culture of the cells. The method reduces the risk of clinical applications of iPSCs by using a combination of highly-safe reprogramming factors without the introduction of high-risk reprogramming factors such as c-MYC, SV40-LT and TP53 inhibitors; The method is highly applicable.

A selection method of an iPS cell includes: at a reprogramming process to culture a cell including a plurality of combinations of initializing factors labelled with luminescent genes that are different with each other, acquiring a photon number per unit area or a photon number per unit time of each of the luminescent genes of the cell; judging whether the acquired photon number is more than a threshold that is predetermined for the acquired photon number; and when the acquired photon number is more than the threshold, selecting this cell as an objective cell for a next process.

The invention provides compositions and methods for reprogramming minimal volumes of mononuclear cells. In particular aspects, the invention provides methods and compositions for reprogramming minimal volumes of umbilical cord blood obtained from cord blood segments from cryopreserved cord blood segments.

Methods and composition of induction of pluripotent stem cells and other desired cell types are disclosed. For example, in certain aspects methods for generating essentially vector-free induced pluripotent stem cells are described. Furthermore, the invention provides induced pluripotent stem cells and desired cell types essentially free of exogenous vector elements with the episomal expression vectors to express differentiation programming factors.

Minivectors having defined, non-transient shapes as determined by sequence are provided, along with uses in the targeting delivery of such minivectors to target tissues for preferential gene delivery. Method of designing and making shaped minivectors are also provided.