8results about How to "Improve expression efficiency" patented technology

This invention discloses a method for mRNA circularization and translation based on ribozyme catalysis. The method utilizes a Td ribozyme sequence to perform in vitro mRNA circularization, yielding circRNA; and introduces an IRES sequence into the circRNA. The IRES sequence is derived from MCDV, CVB3, HCV, PTV-1, RSV, or TSV. Through screening, this invention ultimately determined that the Td ribozyme sequence has the highest circularization efficiency, and that the IRES sequences derived from MCDV, CVB3, HCV, PTV-1, RSV, or TSV have strong ribosome binding capabilities. The MCDV IRES sequence is adaptable to various cellular environments. This invention is of great significance for improving the stability and expression efficiency of mRNA in vivo and promoting the development of circular mRNA technology.

The invention belongs to the field of biological medicine, and particularly relates to a 5 'UTR sequence and application thereof. The technical problem to be solved by the invention is to improve the expression efficiency of mRNA drugs. The main technical scheme of the invention is as follows: two 5 'UTR sequences, which are SEQ ID NO: 1 and SEQ ID NO: 2, are provided; and a nucleic acid molecule comprising the 5 'UTR sequence. Compared with a 5 'UTR sequence optimized in the nature, the mRNA constructed by the 5' UTR has the advantages that the expression efficiency of a target gene is obviously improved; the preparation method is beneficial to reducing the administration dosage and administration frequency of the mRNA drug, reducing the cost of the mRNA drug and reducing the risk of adverse reaction after administration, and has a good application prospect in the industrial production of the mRNA drug.

This invention provides a method for site-specific replacement of the creatine kinase promoter in the Huaxi bovine gene promoter, belonging to the field of animal gene editing and genetic breeding technology. It also provides a method for enhancing the function of bovine muscle cells. FTO Gene expression reagents include crRNA for upstream site knockout, crRNA for downstream site knockout, a Cas12i nuclease expression vector, and an FTO homologous substitution vector; the FTO homologous substitution vector includes... FTO upstream homologous arm of gene promoter, MCK gene promoter and FTO Downstream homologous arm of the gene promoter. The reagent was used in West China bovine cells. FTO Application in gene promoter replacement. This invention can specifically improve the expression efficiency of genes related to fat development in muscle tissue, providing an efficient and precise technical means for the genetic improvement of the performance of beef from Western China.

The present invention relates to an o / w emulsion comprising an oil phase, an aqueous phase and particles dispersed in the aqueous phase, wherein the particles are lipid nanoparticles (LNP) comprising a nucleic acid component, the particles being capable of stabilizing the oil-water interface. The invention also discloses a pharmaceutical composition, such as a vaccine, comprising the o / w emulsion.

This invention provides a high-throughput and efficient method for screening multiple target genes that can regulate mRNA expression levels in cells by co-transfecting or sequentially transfecting mRNA with a gene silencing library targeting genes in cells using lipid nanoparticles. Several target genes that can regulate mRNA protein expression levels through the action of siRNA have been discovered. By manipulating these genes that regulate mRNA expression levels, the translational capacity of mRNA in mRNA vaccine and drug development can be improved, mRNA protein expression efficiency can be increased, and immunogenicity and toxicity can be reduced.

The application provides a repeated protein expression method and integrated device using anti-degradation circular DNA and circular RNA. The repeated protein expression method comprises a DNA template capable of producing circRNA in a cell-free protein expression system; the DNA template is a double-stranded DNA template with closed ends formed by two single-stranded DNAs; the double-stranded DNA template is connected with a closed linker at two ends of a linear DNA template; the closed linker comprises a 5' end linker and a 3' end linker; the sequence of the non-coding region of the 5' end linker and the sequence of the non-coding region of the 3' end linker can be transcribed into a cleavage site of a circRNA precursor, and a circRNA is formed through a sequence complementary affinity effect. According to the technical scheme of the application, the circRNA can be continuously produced in the cell-free protein expression, the degradation of the DNA fragment is solved, the protein expression efficiency is improved, and the template can be repeatedly used after the hydrogel is covalently bonded with the template, so that the production cost is further reduced.

This invention relates to the fields of molecular biology and biotechnology, and provides a reporter vector for visualizing small RNA-target gene regulation. Using a pCOX2 vector as a backbone, it carries a CaMV 35S promoter, a RUBY reporter gene, and the target site sequence of a small RNA target gene, forming a cis-35S-RUBY-target site expression cassette. The CaMV 35S promoter is effectively linked to and drives the expression of the RUBY reporter gene, and the target site sequence is located downstream of the RUBY reporter gene sequence. This invention also provides a method for constructing and applying the aforementioned reporter vector. By constructing a dedicated reporter vector carrying the RUBY reporter gene and combining it with the target site sequence of a small RNA target gene, this invention achieves visual verification of the regulatory effects between small RNA and target genes, offering advantages such as ease of operation, intuitiveness, high efficiency, non-invasiveness, and low cost.

The present application belongs to the technical field of molecular biology, and relates to a truncated fish peptidoglycan recognition protein and application thereof. The amino acid sequence of the truncated fish peptidoglycan recognition protein is shown in Table SEQ ID NO. 1. The truncated fish peptidoglycan recognition protein provided by the present application has higher expression efficiency and higher yield in a prokaryotic expression system compared with a natural peptidoglycan recognition protein. The truncated fish peptidoglycan recognition protein has stronger binding capacity with peptidoglycan (PGN) and lipopolysaccharide (LPS), which are main components of cell walls of gram-positive bacteria and gram-negative bacteria. The bacteriostatic and agglutination capacity of the truncated fish peptidoglycan recognition protein on gram-positive bacteria and gram-negative bacteria is significantly improved. These characteristics show that the truncated fish peptidoglycan recognition protein has development potential as an aquatic immunopotentiator, and provides important support for subsequent application research.