Abstract: Methods for inserting a polynucleotide sequence into the genome of a human cell are provided. The present methods result in insertion of a polynucleotide sequence of interest into the H11 locus in the genome of a human cell. Also provided are nucleic acids that include sequences for integrating a polynucleotide sequence of interest into the H11 locus in the genome of a human cell. A transgenic human cell including site specific recombination sites at the H11 locus is also disclosed.

Abstract: Methods for inserting a polynucleotide sequence into the genome of a human cell are provided. The present methods result in insertion of a polynucleotide sequence of interest into the H11 locus in the genome of a human cell. Also provided are nucleic acids that include sequences for integrating a polynucleotide sequence of interest into the H11 locus in the genome of a human cell. A transgenic human cell including site specific recombination sites at the H11 locus is also disclosed.

Abstract: The present invention describes methods of identifying altered recombinases and compositions thereof, wherein at least one amino acid is different from a parent, wild-type recombinase and the altered recombinase has improved recombination efficiency towards wild-type and/or pseudo att site sequences relative to the parent, wild-type recombinase. The present invention also includes methods of modifying the genomes of cells using the altered recombinases, including methods of site-specifically integrating a polynucleotide sequence of interest in a genome of a eucaryotic cell.

Abstract: The present invention provides methods of site-specifically integrating a polynucleotide sequence of interest in a genome of a eucaryotic cell, as well as, enzymes, polypeptides, and a variety of vector constructs useful therefore. In the method, a targeting construct comprises, for example, (i) a first recombination site and a polynucleotide sequence of interest, and (ii) a site-specific recombinase, which are introduced into the cell. The genome of the cell comprises a second recombination site. Recombination between the first and second recombination sites is facilitated by the site-specific recombinase. The invention describes compositions, vectors, and methods of use thereof, for the generation of transgenic cells, tissues, plants, and animals. The compositions, vectors, and methods of the present invention are also useful in gene therapy techniques.

Abstract: The present invention describes an expression vector useful for transfection of a selected mammalian host cell. The vector includes the following components: an Epstein Barr Virus (EBV) family of repeats; a copy of the EBV Nuclear Antigen-1 (EBNA-1) gene that can be functionally expressed in the host cell; a eucaryotic DNA fragment, which provides the ability of the vector to replicate in host cells; and an expression cassette which comprises a promoter functional in said host cell, a coding sequence having 5' and 3' ends, where said coding sequence is functionally linked to said promoter, where said 5' end is adjacent the promoter and said 3' end is adjacent transcription termination sequences. The vector of the present invention is useful in the transfection of mammalian cells, including rodent and human cells. The vector is stably retained and replicates in concert with genomic sequences of the host cell, that is, the vector is typically replicated once per cell cycle.