11 results about "Bicistronic mrna" patented technology

This invention discloses a recombinant vector, recombinant bacteria, and fermentation method for high-yield GHK production, belonging to the field of biomedical technology. The recombinant expression vector of this application comprises a bicistronic expression cassette controlled by a T7 / lac heterozygous promoter, sequentially containing the encoding (GHK). n The vector encodes the first cistron of the polypeptide and the second cistron, where n is an integer from 2 to 200. When this vector is transformed into *E. coli* BL21, the resulting recombinant bacteria can simultaneously express (GHK) after IPTG induction. n Polypeptides and trypsinogen. During fermentation, trypsinogen is activated and enables the processing of (GHK). n In situ enzymatic cleavage of peptides efficiently releases GHK. This invention integrates the traditional multi-step process into a "one-step fermentation," simplifying the production process and reducing purification difficulty. Examples show that the GHK yield can reach up to 3.9 g / L, providing an efficient and convenient new strategy for the large-scale biomanufacturing of GHK.

This disclosure relates to bicistronic polypeptide constructs for use in allogeneic gene therapy, such as CAR-T cell therapy. A bicistronic construct comprises a first polynucleotide encoding a therapeutic molecule (e.g., CAR-T or antibody) and a second polynucleotide encoding an immune surveillance masking molecule (ISMM). The ISMM includes a non-functional version of a protein knockout, such as a human leukocyte antigen E genetically fused to a fragment, by insertion of a bicistronic construct, such as beta-2 microglobulin or B2M. Also provided are vectors comprising bicistronic constructs, cells (e.g., CAR-T cells), and methods of use. Kits and manufacturing articles are also provided. This disclosure also provides four novel insertion sites that may be used to insert expression constructs into the B2M gene.

This invention relates to a mouse model of Parkinson's disease and its applications. The construction method includes the following steps: 1) in mice... Snack An exogenous fragment containing the human SNCA G51D coding sequence was knocked into the region upstream of the gene stop codon via homologous recombination, causing the human SNCA G51D coding sequence to interact with the mouse endogenous gene. Snack The gene forms a bicistronic expression structure, and the coding sequence of the human SNCA G51D is shown in SEQ ID No:1; 2) Positive F0 generation mice are backcrossed with wild-type mice, and genotypes are identified and screened to obtain a stable SNCA G51D endogenous knock-in Parkinson's disease mouse model. The gene editing strategy of the Parkinson's disease mouse model preserves the mouse's... Snack Simultaneous endogenous expression of α-synuclein G51D in the gene can mimic two pathogenic forms, exhibiting high physiological relevance and genetic authenticity.

The present disclosure provides nucleic acid molecules (e.g., a plasmid or vector) comprising a nucleic acid sequence encoding a bicistronic or multicistronic LAMP Construct comprising specific fragments of the LAMP luminal domain and an antigenic domain heterologous to the LAMP protein to provide at least one antigen for priming an immune response, wherein the antigen expressed by the Construct is optionally processed and presented to MHC class II molecules, and also a nucleic acid sequence encoding an immune response enhancing polypeptide that is optionally secreted from a host cell. The nucleic acid molecules can be used, for example, for the treatment of disease and in particular, allergies, infectious disease, diabetes, hyperproliferative disorders and / or cancer.

This invention provides a bicistronic CAR-T cell structure that simultaneously targets CD19 and CD72 and its application. The nucleic acid molecule encoding the corresponding chimeric antigen receptors includes at least nucleotide sequences encoding a first chimeric antigen receptor, a second chimeric antigen receptor, and a cleavage peptide. The nucleotide sequence encoding the cleavage peptide is located between the nucleotide sequences encoding the first and second chimeric antigen receptors. The first antigen-binding domain is specific for CD19, and the second antigen-binding domain is specific for CD72. This nucleic acid molecule is a bicistronic structure capable of independently expressing both CD19 and CD72, achieving balanced co-expression of these two independent CAR-T molecules (CD19 CAR and CD72 CAR), thus providing an effective treatment option for relapsed / refractory patients.

This specification provides methods for detecting the expression of bicistronically expressed polypeptides, for example, in a population of immune cells, such as a population of CAR T cells, that have been manipulated into chimeric cytokine receptors. In one aspect, this disclosure provides an in vitro method for detecting bicistronically expressed polypeptides in chimeric antigen receptor (CAR) T cells.

Embodiments described herein provide orthohepadnavirus capsid protein (Cp) heterodimers, bicistronic vectors encoding the heterodimers, and methods for producing the heterodimers. The heterodimers can be used to form mosaic virus-like particles. In certain embodiments, the heterodimers can form a hexamer, which in turn can be used to nucleate capsid formation, resulting in a Janus particle-like virus-like particle. The hexamer's can then be removed, leaving holey capsids. The capsids can be loaded with, for example, one or more polypeptides, small molecules, or a combination of polypeptides and small molecules. The holes of the holey capsids can be filled with another orthohepadnavirus heterodimer or a homodimer.

The present invention relates to a screening composition for a therapeutic agent for coronavirus infection, comprising a CoV RdRp expression vector and a bicistronic reporter vector, a screening kit for a therapeutic agent for coronavirus infection, comprising the composition, and a method for screening a therapeutic agent for coronavirus infection using the composition or kit. When the screening composition for a therapeutic agent for coronavirus infection, provided by the present invention, is used, candidate materials that can have direct influences on the activity of CoV RdRp can be screened more quickly and easily, and thus, the composition can be widely used in the development of therapeutic agents for coronavirus infection.

This invention discloses a bicistronic translation-coupled expression vector. The expression vector is constructed by inserting a molecular chaperone, a novel ribosome binding site, and the target protein gene into the pET-28a(+) plasmid. Furthermore, this invention constructs a genetically engineered bacterium using *E. coli* BL21(DE3) as the expression host. Utilizing the bicistronic translation-coupled expression vector, the co-expression of mussel protein Mgfp-3B with the molecular chaperones SUMO and / or TrxA is achieved, promoting the correct folding and translation of Mgfp-3B for expression in a soluble form. This invention enables a simple and rapid production of bioactive soluble mussel protein in *E. coli*, and the bicistronic expression vector avoids the enzymatic chaperone manipulation required for fusion expression. In addition, this invention, through optimized fermentation medium and fermentation process, achieves a soluble mussel protein yield of 200-300 mg / L, simplifying downstream purification processes.

Aspects of the disclosure relate to bicistronic AAV nucleic acid constructs comprising a transgene encoding hexosaminidase A (HEXA) and hexosaminidase (HEXB) proteins. In some embodiments, the disclosure provides methods for treating or preventing lysosomal storage disorders, such as Tay-Sachs disease and Sandhoff disease, using bicistronic nucleic acid constructs described by the disclosure.

The invention discloses a recombinant vector for high yield of GHK, recombinant bacteria and a fermentation method, and belongs to the technical field of biological medicines. The recombinant expression vector comprises a bicistronic expression cassette controlled by a T7 / lac hybrid promoter, the bicistronic expression cassette sequentially comprises a first cistronic for coding (GHK) n polypeptide and a second cistronic for coding trypsinogen, and n is an integer from 2 to 200. The vector is transferred into Escherichia coli BL21, and the obtained recombinant bacterium can synchronously express the GHK n polypeptide and trypsinogen after being induced by IPTG (isopropyl-beta-d-thiogalactoside). In the fermentation process, trypsinogen is activated and in-situ enzyme digestion of the GHK n polypeptide is realized, so that the GHK is efficiently released. According to the method, a traditional multi-step process is integrated into one-step fermentation, the production process is simplified, the purification difficulty is reduced, the highest GHK yield can reach 3.9 g / L according to the embodiment, and an efficient and convenient new strategy is provided for large-scale biological manufacturing of GHK.