The present invention provides a nucleic acid sequence comprising at least one miRNA binding site sequence containing at least one miRNA binding site. Those miRNA binding site sequences are located within and/or immediately 3 or 5 of the 5 UTR of a gene to reduce the off-target side effects and allow a cell type specific expression from the nucleic acid sequence within the target organ or organs. The invention further provides pharmaceutical compositions, as well as a method of promoting cell-type specific expression, comprising the nucleic acid sequence according to the invention for use in therapy.

Nucleic acid molecules comprising a coding sequence with at least one codon substituted to a synonymous codon, a modified form of a virus comprising the nucleic acid molecules of the invention, and methods for producing these nucleic acid molecules, and viruses, are provided.

The present disclosure describes methods and systems for use in the production of recombinant adeno-associated virus (rAAV) particles. In certain embodiments, the production process and system include engineered untranslated regions (UTR) which allow for the controlled expression of AAV capsid proteins, such as VP1, VP2 and VP3. In certain embodiments, the production process and system include engineered untranslated regions (UTR) which allow for the controlled expression of non-structural AAV replication proteins, such as Rep78 and Rep52.

A nucleic acid molecule can code for an Orf virus vector promoter. A recombinant Orf virus vector can be included in a cell. The nucleic acid molecule, the vector and/or the cell can be included in a composition. The recombinant Orf virus vector can be used for the production of a foreign gene.

Disclosed herein include methods, compositions, and kits enabling expression of multiple payload proteins from a single mRNA configured to achieve a predetermined stoichiometry of first unit payload protein(s) and second unit payload protein(s) in a cell or cell-like environment. There are provided, in some embodiments, nucleic acid compositions comprising a polynucleotide comprising a first nucleic acid unit (encoding one or more first unit payload protein(s)) and a second nucleic acid unit (encoding one or more second unit payload protein(s)). The first nucleic acid unit and the second nucleic acid unit can each comprise an engineered translation initiation site (eTIS) comprising a three-nucleotide tunable element immediately upstream of a start codon. In some embodiments, the nucleic acid composition comprises at least one modified nucleotide and/or at least one nucleotide analogue or nucleotide derivative. In some embodiments, the polynucleotide comprises a disclosed eTIS combination.

The present disclosure relates to a nucleic acid molecule including plural translation control elements downstream inserted downstream of a coding region, and a nucleic acid expression platform or a nucleic acid expression system such as a recombinant expression vector including the nucleic acid molecule inserted therein. A target gene inserted into the coding region can be expressed efficiently using the nucleic acid molecule. It is possible to improve expression efficiency of a report gene and, an antigen and/or therapeutic protein or peptide utilizing the expression platform.

The present invention relates to recombinant DNA molecules such as plasmids, non-viral vectors, viral vectors and hybrids thereof, and more particularly to vector modifications that improve expression of said DNA molecules in cell lines and organisms.

Materials and methods for genome engineering through transient expression of a targeted nuclease are described herein. For example, the methods described herein can include introducing into a cell a messenger RNA (mRNA) that encodes a nuclease targeted to a selected sequence within the cell, where the stability of the mRNA is modified by the addition of untranslated regions (UTRs).

The present invention relates to a gene expression cassette including a synthetic 5 untranslated region (5 UTR), a promoter, and a regulatory gene; a recombinant vector including a replication origin and the gene expression cassette; a recombinant microorganism which has the recombinant vector introduced thereinto and shows alleviated segregational instability and; a method for preparing a recombinant microorganism having alleviated segregational instability by introducing the recombinant vector thereinto; and a method for quantitatively controlling a plasmid copy number in a recombinant microorganism. According to the present invention, removal of infA and efp, which are genes indispensable for cells, encoding respectively for a translation initiation factor and a protein elongation factor (EF-P), from a microbial chromosome and introduction of the gene expression cassette including the regulatory gene with Escherichia coli serving as a host allow the stable maintenance of plasmids in an antibiotic-free medium without causing intercellular intrinsic variations. In addition, the precise control of expression levels of infA and efp in the recombinant microorganism by means of a promoter can lead to the quantitative control of PCN at high yield as well. Therefore, the present invention can find a broad spectrum of applications in a variety of industries producing recombinant proteins.

The technology relates to a nucleic acid expression cassette comprising a TR element encoding an mRNA molecule that is translated in stressed and/or dying cells, and a nucleotide sequence operably linked to the TR element, that is a first open reading frame (ORF) sequence and encodes a polypeptide or a fragment thereof and is co-translated with the TR element. The technology further relates to mammalian cells and a transgenic animal comprising such expression cassette. Further included are kits comprising the expression cassette, and methods for determining toxicity, and killing a target cell.