25 results about "Nuclear localization sequence" patented technology

The invention relates to a gene editing delivery vector, a delivery system and a preparation method and application thereof. The carrier is a cell nucleus targeting polymer carrier prepared by modifying linear polyethyleneimine with phenylboronic acid. Nanoparticles and a nanometer delivery system prepared by using the vector can have ultrahigh cell nucleus targeting ability without additional traditional nuclear localization sequence (NLS) peptide modification, can perform efficient gene editing in living cells and in vivo levels, and can perform real-time imaging on target genes in a cell nucleus region, so that the target genes can be efficiently edited. And the efficiency of living cell and in-vivo gene editing and the accuracy of nucleic acid detection are remarkably improved. Meanwhile, the kit has the advantages of easiness in preparation, high gene editing efficiency, high detection sensitivity and the like.

The invention discloses a probe set. The probe set comprises a first fusion protein and a second fusion protein, wherein the first fusion protein comprises a nuclear localization sequence and a first protein molecule, and the nuclear localization sequence is connected with the first protein molecule; the second fusion protein comprises a TFEB protein and a second protein molecule, the TFEB protein is connected with the second protein molecule, the second protein molecule is used for being combined with the first protein molecule, and a detectable signal is generated after the first protein molecule and the second protein molecule are combined. The probe set can rapidly, accurately and sensitively detect the TFEB nuclear transfer condition, can be used for high-throughput screening of TFEB regulation and control drugs, and reduces the drug screening cost.

This invention discloses a protease-responsive GPCR receptor and its applications. The protease-responsive GPCR receptor of this invention comprises an extracellular protease recognition domain, a transmembrane domain, and an intracellular signaling domain; the extracellular protease recognition domain contains a peptide hydrolyzed by the protease; the intracellular signaling domain contains an HCV enzyme hydrolyzed peptide, a nuclear localization sequence (NLS), a Cas protease, and a coupled nuclear transcription factor. This protease-responsive GPCR receptor can induce endogenous dCas9 assembly into the nucleus and activate the chimeric antigen receptor signaling pathway in natural killer cells in response to extracellular proteases, providing a new solution for developing novel precision immunotherapy.

The present invention provides for a synthetic transcription factor (TF) comprising (a) a DNA-binding domain of a transcription factor linked to (b) an activator domain (AD), and (c) optionally a nuclear localization sequence (NLS). The present invention provides for a nucleic acid encoding an activator domain (AD) of the present invention.

The present invention belongs to the field of biomolecular technology, and in particular to a kind of RNA-modified live cell imaging system and its composition and application. The RNA-modified live cell imaging system comprises an RNA reporter system and a FRET probe, wherein the RNA reporter system comprises a tetracycline-induced expression system, an mMaroon1 fluorescent protein, an NLS-nuclear localization sequence and a 22-repeat m6A-MS2 reporter sequence; the FRET imaging probe comprises a YTH domain, a YPet fluorescent protein, a long-chain flexible linker, an ECFP fluorescent protein, an MCP protein and an NLS nuclear localization signal. The m6A-modified live cell imaging system developed by this patent can perform high-resolution real-time dynamic imaging of m6A modification in living cells, and can realize triple dynamic monitoring of m6A modification levels and RNA localization and translation, thereby building a research platform and drug screening platform for m6A modification pathways.

The invention discloses a USAG-1 recombinant protein targeted delivery system based on multi-enzyme logic gating and a preparation method, and mainly relates to crossing of biological medicine and nanotechnology. Comprising the following steps: S1, a C-terminal fusion nuclear localization sequence PKKKRKV of a recombinant USAG-1 protein expressed by CHO cells, and the purity of the recombinant USAG-1 protein is greater than or equal to 98%; s2, preparing core-shell structure nanoparticles, wherein the core is PLGA loaded BMP-2, and the drug loading capacity is 2.5-4.2 [mu] g / mg; the outer-layer lipid membrane is composed of DSPC / DSPG according to the ratio of 7: 3, and the surface embedded USAG-1 molecular density is greater than or equal to 1 * 10 / mu m < 2 >; s3, the surface co-modified EGFR antibody and the RGDfK peptide are connected through a thiol-maleimide bond and a PEG2000 spacer arm according to the molecular ratio of 1: 1.6 + / -0.2, the coding sequence of the recombinant USAG-1 is regulated and controlled by an MMP-9 response promoter, and the promoter is located in MMP-9gt; and the expression is activated at 20nM. The system has the beneficial effects that time-space precise delivery of the bone metabolism related protein USAG-1 is realized through a multi-enzyme logic gating mechanism, and the system is suitable for treatment of bone metabolism disorder diseases such as osteosarcoma and alveolar bone defect.

The invention discloses a shuttle peptide targeting YY1S247 phosphorylation and an application of the shuttle peptide. The sequence of the shuttle peptide is as shown in SEQ ID NO: 1. Wherein the RQIKIWFQNRRMKWKK is a part of the drosophila melanogaster contact foot peptide and is used for providing cell penetrability; the PKKKRKV is a nuclear localization sequence of the SV40 large T antigen and is used for guiding a peptide fragment to enter a cell nucleus; and the QIIGENSPPDYSE is a peripheral sequence of the YY1S247 site. Results show that the shuttling peptide (CPP) can competitively, specifically and effectively inhibit phosphorylation of the YY1S247 site, so that CD24 expression is inhibited, and the treatment effect of EGFR-TKI is enhanced.

The invention relates to the field of bioengineering, in particular to an expression cassette set, a telomere-telomerase system and application of the telomere-telomerase system. The invention provides an expression cassette group. The expression cassette group comprises an expression cassette 1 and an expression cassette 2, the expression cassette 1 comprises a URA3 tag, an ACT1 gene and a tos sequence; the expression box 2 comprises a telN gene and a nuclear localization sequence. The telomere-telomerase system provided by the invention can control formation and loss of linear chromosomes. And recognizing and cutting the tos site by utilizing a TelN enzyme to form a linear chromosome with a hairpin closed tail end. Meanwhile, the existence of the TelN enzyme can enable the linear chromosome to be stably copied, so that the formation and degradation of the linear chromosome can be controlled by controlling the expression of the TelN enzyme.

Disclosed are a novel nuclear localization sequence (NLS) mutant and a method for improving biosynthetic efficiency using same, belonging to the field of bioengineering. According to the present disclosure, a novel NLS is screened out, and recombinant yeast with improved metabolite synthesis efficiency is constructed by applying the NLS obtained through screening, which provides key technical support for developing a high-yield, stable and cost-effective yeast biosynthesis platform, and facilitates the use of recombinant yeast as a cell factory for large-scale industrial biomanufacturing.

The application relates to the technical field of biological medicine engineering, and provides a preparation method and application of a scorpion toxin-derived transmembrane polypeptide. The OpiCalcin1 polypeptide is optimized in a truncation mode, two polypeptides Opi1 17-33 CSKKCKRRGTNPEKRCR and Opi1 23-33 RRGTNPEKRCR with excellent transmembrane function and low cytotoxicity are provided. Through experiment verification, the two cell transmembrane peptides have excellent transmembrane effect and low cytotoxicity, meanwhile, the defects of time-consuming and laborious in synthesizing the complete sequence of scorpion toxin OpiCalcin1 are overcome, the cell transmembrane peptides can be used for preparing drug, nuclear localization sequence and probe transport carriers, and the targeting efficiency is improved. Compared with traditional scorpion toxin medicine, the synthesis of the scorpion toxin to explore the transmembrane activity provides a new selection for the drug carrier.

In one aspect, the invention relates to a TnpB variant polypeptide comprising a Deinococcus radiodurans (ISDra2) TnpB polypeptide sequence and, separated by a peptide linker, a nuclear localization module comprising at least two nuclear localization sequences (NLS) separated by a first peptide linker. A second aspect of the invention relates to a DNA editing polypeptide comprising the TnpB variant polypeptide according to the invention, wherein the TnpB variant polypeptide comprises a nuclease deactivating mutation, and a nucleoside deaminase. The invention further relates to a nucleic acid sequence encoding a polypeptide according to the invention, and to a virus comprising the nucleic sequence.

The present invention belongs to the field of biomedicine, and specifically relates to a gene delivery method based on exosomes and polypeptides containing nuclear localization sequences. Specifically, the present invention provides a method for increasing the expression of exogenous genes in target cells through exosomes, the method comprising the steps of mixing exosomes with exogenous genes and co-electroporating them with cells; the method also comprises mixing and incubating the exogenous genes with auxiliary polypeptides and / or KK58 before electroporation, wherein the auxiliary polypeptides are composed of nuclear localization signals, nuclear localization signals and DNA binding domains.

The application discloses a CRISPR / Cas9-drug co-assembly biomimetic nano system and preparation and application thereof, and belongs to the technical field of biological medicines. The biomimetic nano system is composed of a natural product active small molecule drug as a nano inner core, a gene editing system and an auxiliary component co-assembled into a nano drug particle, and then a biomimetic membrane is further wrapped on the surface to form the biomimetic nano system; wherein the natural active small molecule drug is ursolic acid, the CRISPR / Cas9 gene editing system is a ribonucleoprotein complex containing a nuclear localization sequence, the auxiliary component is a cell-penetrating peptide, and the biomimetic membrane is a tumor cell membrane. The biomimetic nano system is constructed on the basis of revealing the combined immune regulation characteristics and molecular mechanisms of immune checkpoint PD-L1 gene therapy and natural product ursolic acid anti-tumor, has a significant inhibitory effect on the occurrence of liver cancer, and has a wide application prospect in the preparation of anti-liver cancer drugs.

The present technology relates to the field of drug delivery and provides a molecule comprising or consisting of at least one protein-based carrier building unit, wherein the protein-based carrier building unit comprises at least one, preferably at least two attachment points or conjugation sites. In particular, the present technology provides a molecule comprising at least one protein-based building unit, wherein the at least one protein-based building unit: a) comprises at least one conjugation site or attachment point; b) has a molecular weight of 2.5 to 70 kDa; c) has a globular three-dimensional (3D) structure; d) has a solubility of 10 mg / mL or more, measured in aqueous solution at room temperature; and does not specifically bind to any human protein or binds to one or more human proteins with a K D value of more than 5 x 10 ‑4 mol / liter, wherein the molecule further comprises at least one nuclear localization sequence (NLS) covalently linked to the at least one conjugation site or attachment point comprised in the protein-based carrier building unit, directly or via a linker, wherein the NLS preferably comprises or consists of SEQ ID NO: 221.

This invention discloses a shuttle peptide targeting YY1S247 phosphorylation and its application. The sequence of the shuttle peptide is shown in SEQ ID NO: 1. RQIKIWFQNRRMKWKK is a part of the Drosophila melanogaster tentacles peptide, used to provide cell penetration; PKKKRKV is the nuclear localization sequence of the SV40 large T antigen, used to guide the peptide into the cell nucleus; QIIGENSPPDYSE is the surrounding sequence of the YY1S247 site. The results show that the shuttle peptide (CPP) of this invention can competitively and specifically inhibit phosphorylation at the YY1S247 site, thereby inhibiting CD24 expression and enhancing the therapeutic effect of EGFR-TKIs.

A gene editing system of Candida viswanathii includes a Candida viswanathii, a first gene editing fragment and a second gene editing fragment. The first gene editing fragment successively includes a first homology arm and a screening gene. The second gene editing fragment is connected to a C-terminus of the first gene editing fragment and includes a second homology arm, a Cas9 expression cassette and a sgRNA cassette. The Cas9 expression cassette successively includes a Cas9 promoter, a Cas9 gene and three nuclear localization sequences. The sgRNA cassette successively includes a sgRNA promoter, a first ribozyme, a targeting sequence, a scaffold and a second ribozyme. The first gene editing fragment and the second gene editing fragment are constructed as a linear fragment for gene editing of a chromosome of the Candida viswanathii.

The invention discloses a novel nuclear localization sequence mutant and a method for improving biosynthesis efficiency thereof, and belongs to the field of bioengineering. According to the invention, a new nuclear localization sequence is screened, and the recombinant yeast with improved metabolite synthesis efficiency is constructed by using the nuclear localization sequence obtained by screening, so that a key technical support is provided for developing a high-yield, stable and economic yeast biosynthesis platform; and large-scale industrial biological manufacturing by using the recombinant yeast as a cell factory is facilitated.

The present invention relates to a gene expression regulation optimization technology based on a target DNA recognition ultra-small molecular module, and relates to an ultra-small eCWCas12f-VPR system optimized through engineering of a CRISPR-Cas module, linking of activation domains, and various combinations of linkers and NLS sequences. It has been identified that effective and target-specific gene expression regulation superior to that of a conventional CRISPRa system can be achieved and the accuracy of the gene expression regulation is remarkably excellent. In particular, unlike a conventional CRISPR gene scissors technology, the system of the present invention can accurately regulate the expression of a target gene without cutting or damaging DNA, and thus is relatively safe and optimized for application in a gene therapy method, thereby exerting a great impact, and, due to the characteristics of ultra-small size, enables all genes to be loaded in a single vector (all-in-one), and thus can be effectively used as an excellent novel gene editing technology that compensates for the disadvantages of a conventional gene editing technology.

The present description relates to a conjugated compound an antibody covalently linked to enhancer moiety composed by a nuclear localization sequence (NLS), covalently linked to a sterol variant, such as cholic acid (ChAc) or a variant thereof. The enhancer moiety as encompassed herein is able to induce endosome escape of the compound-conjugates by direct membrane destabilization or indirectly by ROS and ceramide production which destabilize endosome-lysosome membrane. The conjugated compound can further comprise payload.

The invention features polypeptides containing nuclear localization sequences that can be used to deliver polynucleotides to a cell. The polypeptides can be formulated with a polynucleotide and a lipid.

An engineered phage-derived particle (PDP) for expressing a transgene in a target cell transduced with a bacteriophage, the PDP includes (i) less than about 500 bp of DNA from the bacteriophage genome, (ii) an ITR-flanked therapeutic gene up to 20 kb, (iii) an endosomal escape sequence, (iv) a nuclear localization sequence, and (v) a cell-specific targeting moiety. The PDP may escape lysosomal degradation, traffic across the nuclear envelope and expressed a therapeutic gene in a mammalian cell.

The present technology relates to the field of drug delivery and provides molecules comprising or consisting of at least one protein-based carrier building block, wherein the protein-based carrier building block comprises at least one, preferably at least two, attachment point(s) or conjugation site(s). In particular, the technology provides a molecule comprising at least one protein-based building block, wherein the at least one protein-based building block: a) comprises at least one conjugation site or attachment point; b) has a molecular mass of 2.5 to 70 kDa; c) has a globular three-dimensional (3D) structure; d) has a solubility of 10 mg / mL or more, measured in an aqueous solution at room temperature; and does not specifically bind to any human protein or binds one or more human proteins with a KD value greater than 5x10-4 mol / litre, wherein the molecule further comprises at least one nuclear localization sequence (NLS), covalently linked, directly or by means of a linker, to at least one conjugation site or attachment point comprised in the protein-based carrier building block, wherein the NLS preferably comprises or consists of SEQ ID NO.: 221.

Retron editors allow for the insertion of a segment of user-defined DNA in a host cell genome. To maximize the activity of retron editors, they were optimized by improved linkers, nuclear localization sequences, and RNA sequences. The resulting retron editor can programmably insert, delete, or change the genome of a host cell or organism, including human genomes.

The present invention relates to the field of gene transfer, in particular, targeted gene transfer and enzymes for use in targeted gene transfer. In particular, the present invention provides a nucleic acid encoding a fusion protein having serine integrase activity, wherein the fusion protein comprises a serine integrase consisting of a protein having at least 80% sequence identity to the large serine integrase Pa01 and a nuclear localization sequence at the C-terminus of the serine integrase. The invention further provides a fusion protein encoded by said nucleic acid as well as methods of preparing an engineered cell using the nucleic acids and / or fusion proteins of the invention. Finally, the present invention provides various pharmaceutical compositions and their use in treating a disease or disorder.