Millipore Corporation

Cobra Biologics Limited

Emd Millipore Corporation

Cobra Therapeutics Limited

Steven Geraint Williams

Alistair Simpson Irvine

David J Sherratt

Robert L Crombie

Kai S Lipinski

Julian Aj Hanak

Julian A J Hanak

David John Simpson

Scott D Gorman

Jonathan Gawn

Julian Alexis John Hanak

The invention relates to improved genetic elements providing high levels of expression of operably-linked genes in a variety of tissues. In particular, fragments of unmethylated, CpG islands of less than 2 kb are shown to provide enhanced transgene expression and have advantages in terms of vector construction and cloning capacity.

Expression elements

Polynucleotides and vectors comprising an expressible nucleic acid flanked by a 5' extended methylation-free CpG island and a 3&#x2032; selectable marker element are disclosed. Such polynucleotides and vectors provide a means for obtaining high levels of expression of the flanked expressible nucleic acid. Preferred embodiments include combinations of 5&#x2032; extended methylation-free CpG islands and 3&#x2032; antibiotic resistance genes.

Vectors comprising CpG islands without position effect varigation and having increased expression

The invention relates to genetic elements capable of improving the levels of expression of operably-linked transcription units. In particular, said genetic elements are derived from the 5' untranslated regions of ribosomal protein genes and may comprise a CpG island. Also provided are vectors and host cells comprising said genetic elements and methods of use to obtain high levels of recombinant gene expression.

The invention comprises novel polynucleotides, and related vectors, host cells, methods, and compositions, containing transcriptional enhancers providing very high levels of expression of operably-linked expressible nucleic acid sequences in eukaryotic cells. Advantageously the enhancers may be used in combination with their naturally-associated promoters and/or other genetic elements that increase transcription. The invention comprises eukaryotic expression vectors that are capable of providing increased levels of expression in many cell types over that obtainable from human or murine CMV IE enhancer/promoter elements.

Vectors comprising guinea pig CMV regulatory elements

The invention relates to host cells and methods of preparing a substantially RNA-free cellular component, comprising culturing cells producing the cellular component in a medium and lysing the cells to produce a cell lysate, wherein the cell lysate contains the cellular component and sufficient RNase activity to degrade substantially all of the RNA molecules present in the cell lysate. The invention also relates to substantially RNA-free cellular components.

Purification of cellular components that are substantially RNA free

A system is described which utilizes a novel system of repressor titration for maintenance of a plasmid useful in gene therapy and production of a recombinant protein. The system utilizes a transformed host cell containing a plasmid including an operator susceptible to binding by a repressor expressed in trans, a first chromosomal gene encoding the repressor, and a second chromosomal gene that is functionally associated with an operator and essential for cell growth, wherein the plasmid is present in the cell in sufficient numbers to titrate the repressor such that the essential gene is expressed, thereby permitting cell growth.

Growing community of inventors

Cheshire, United Kingdom

Steven Geraint Williams