The invention relates to nucleic acid modifications for a directed expression modulation by the targeted insertion or removal of CpG dinucleotides. The invention also relates to modified nucleic acids and expression vectors.

The current invention reports a promoter that has the nucleic acid sequence of SEQ ID NO: 02 or SEQ ID NO: 03 which is a human CMV major immediate-early (hCMV-MIE) promoter/enhancer with C to G point mutation at position 41 and/or 179 relative to the transcription start site. This new promoter is especially useful for the production of polypeptides at large scale as it shows reduced promoter silencing and improved polypeptide production.

Provided is an expression vector having an improved ability to express a gene. Also provided are cells transformed by the expression vector and a method for mass-producing a target protein by using the cells. The expression vector contains a simian virus 40 promoter, a scaffold attachment region or matrix attachment region element, and a chimeric intron. The vector shows an improved ability to express a gene, and thus, can attain a significantly increased expression of a heterogeneous gene.

The present disclosure teaches the treatment of a blood pathology, such as a blood pathology associated with impaired hemoglobin synthesis including the treatment of -thalassemia or a related hemoglobinopathy. An RNA molecule such as a short interfering RNA or a hairpin RNA which targets an mRNA species encoding -globin is administered to a subject to reduce the amount of -globin produced to non-zero levels and ameliorate the effects of an - and -globin chain imbalance.

The invention relates to nucleic acid modifications for a directed expression modulation by the targeted insertion or removal of CpG dinucleotides. The invention also relates to modified nucleic acids and expression vectors.

Methods of inducibly targeting a chromatin effector to a genomic locus are provided. Aspects of the methods include employing a chemical inducer of proximity (CIP) system. Aspects of the invention further include methods of screening candidate agents that modulate chromatin-mediated transcription control and methods of inducibly modulating expression of a coding sequence from genomic locus. Also provided are compositions, e.g., cells, reagents and kits, etc., that find use in methods of the invention.

The present invention provides, among other things, mRNA constructs and compositions and methods for reverse gene therapy, including administering to a subject in need of treatment a mRNA gene transfer construct comprising a reverse complement sequence encoding a protein of interest and a sequence encoding a human L1 retro-element.

The present invention solves the problem of providing more efficient barrier insulators to avoid vector silencing and to increase expression in the setting of gene transfer vectors, more particularly in the setting of gene transfer retroviral vectors. In this sense, the authors of the present invention have developed an improved insulator element, namely element IS2, which comprises the following combination of nucleic acid molecules, namely nucleic acid molecule HS4-650 bp as shown in SEQ ID No 2 and a synthetic S/MAR nucleic acid molecule containing 5 M/SARs recognition signatures (MRS) as shown in SEQ ID no 1.

A method of delivering a transgene to a cell is provided. The method uses a vector that contains a T-cell receptor alpha locus control region (TCRLCR) derived gene regulatory cassette having fewer than 5.0-kb. The method delivers the transgene with spatiotemporally specific gene expression and silencing-prevention controls to a cell such that a predetermined subset of progeny cell-types express a gene product from the transgene. Other progeny of the cell diminish, or silence, the expression of the gene product.

The invention provides nucleic acid and polypeptide sequences encoding antibody based scaffolds such as full antibodies, antibody Fab fragments, single chain antibodies, soluble VEGF receptor-Fc fusion proteins, and/or anti-angiogenic PDGF receptors. Also encompassed are cell lines encoding such anti-angiogenic antibody scaffolds, VEGF receptors, and/or PDGF receptors. The invention also provides encapsulated cell therapy devices that are capable of delivering such anti-angiogenic antibody scaffolds, VEGF receptors, and/or PDGF receptors as well as methods of using these devices to deliver the anti-angiogenic antibody scaffolds, VEGF receptors, and/or PDGF receptors to medically treat disorders in patients, including ophthalmic, vascular, inflammatory, and cell proliferation diseases.