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.

Non-conventional yeasts are disclosed herein comprising at least one RNA-guided endonuclease (RGEN) comprising at least one RNA component that does not have a 5′-cap. This uncapped RNA component comprises a sequence complementary to a target site sequence in a chromosome or episome in the yeast. The RGEN can bind to, and optionally cleave, one or both DNA strands at the target site sequence. An example of an RGEN herein is a complex of a Cas9 protein with a guide RNA. A ribozyme is used in certain embodiments to provide an RNA component lacking a 5′-cap. Further disclosed are methods of genetic targeting in non-conventional yeast.

The invention relates generally to methods of generating induced pluripotent stem cells (iPSCs) that do not contain the reprogramming vector. In some embodiments, the invention relates to inducing pluripotency in somatic cells by introducing an episomal vector(s) comprising at least one expression cassette containing reprogramming factors and/or synthetic transcription factors and a suicide gene. In some embodiments, the invention relates to inducing pluripotency in somatic cells by introducing episomal vector(s) comprising expression cassettes containing reprogramming factors and/or synthetic transcription factors and a transcriptionally regulated EBNA-1 gene. In some embodiments, the invention relates to inducing pluripotency in somatic cells by introducing episomal vector(s) comprising expression cassettes containing reprogramming factors and/or synthetic transcription factors and both a suicide gene and a transcriptionally regulated EBNA-1 gene.

A method for treating an HIV disease in a subject in need of said treatment, comprising administering to the subject a therapeutically effective amount of a DNA vaccine comprising an expression vector and a pharmaceutically acceptable excipient, where the expression vector comprises: (a) a heterologous promoter operatively linked to a DNA sequence encoding a nuclear-anchoring protein, where the nuclear-anchoring protein comprises: (i) a DNA binding domain which binds to a specific DNA binding sequence, and (ii) a functional domain of the Bovine Papilloma Virus Type 1 E2 protein, where the functional domain binds to a nuclear component; (b) a multimerized DNA sequence that forms a binding site for the nuclear anchoring protein; and (c) at least one expression cassette comprising a DNA sequence encoding a protein or peptide that stimulates an immune response specific to the protein or peptide; where the expression vector lacks an origin of replication functional in mammalian cells.

Methods of producing hepatocyte lineage cells are provided. The method can include transfecting a cell with one or more expression vectors. For example, a cell can be transfected with a first expression vector containing a first gene that encodes CCAAT/enhancer binding protein alpha (CEBPA), a second expression vector containing a second gene that encodes CCAAT/enhancer binding protein beta (CEBPB), a third expression vector containing a third gene that encodes forkhead box A1 (FOXA1), and a fourth expression vector containing a fourth gene that encodes forkhead box A3 (FOXA3). The method can include culturing the transfected cell obtained in a growth environment. The transfected cell can be cultured in Williams' E medium, ReproFF (feeder-free media maintaining pluripotency) medium, or both. Transfected and/or hepatocyte lineage cells obtained by a method of the present invention are also provided.

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 present invention relates to a polynucleotide comprising at least one promoter and an S/MAR element, wherein said S/MAR element is located downstream of said promoter and wherein the nucleic acid sequence of said S/MAR element (S/MAR sequence) comprises at least 3sequence motifs ATTA (SEQ ID NO:1) per 100 nucleotides over a stretch of at most 200 nucleotides; the present invention further relates to a composition and to a host cell comprising said polynucleotide, and to the polynucleotide for use in medicine and for use in treating genetic disease. The present invention also relates to a kit and to a device comprising said polynucleotide, and to methods and uses related to the polynucleotide.

The invention relates to means for carrying out conjugation between bacteria, and in particular the invention relates to carrier bacteria comprising antimicrobial agents and methods of use. The carrier bacteria are capable of conjugative transfer of DNA encoding the agent to a target cells. The invention further relates to growth or feed conversion ratio promotion in animals. The invention further relates to killing Salmonella or inhibiting the growth or proliferation of Salmonella.

Provided are genetically engineered induced pluripotent stem cells (iPSCs) and derivative cells thereof expressing a chimeric antigen receptor (CAR) and methods of using the same. Also provided are compositions, polypeptides, vectors, and methods of manufacturing.

Provided is: a transgenic plant that has a modified flower color; a self- or cross-fertilized descendant of the transgenic plant; or a propagule, a portion of a plant body, tissue, or cells from the transgenic plant or the self- or cross-fertilized descendant of the transgenic plant. The present invention causes both anthocyanin delphinidin and a flavone C-glycoside that is methylated at the hydroxyl group at the 7 position to be present in the cells of a plant.