42results about How to "Reduce off-target effects" patented technology

The invention relates to the field of biological medicine, in particular to a nucleoside analogue and application thereof to reduction of off-target activity of oligonucleotide drugs. The nucleoside analogue provided by the invention can be used for preparing oligonucleotide drugs, and can be coupled to a seed region of siRNA, so that the specificity of siRNA is efficiently improved, and the silence effect of a target gene can be maintained while the off-target effect of siRNA is reduced.

The invention provides organic nanoparticles with high osteogenic performance as well as a preparation method and application thereof, and belongs to the technical field of osteogenic drug preparation. The preparation method of the organic nanoparticles with high osteogenic performance comprises the following steps: (1) preparing fluorinated polyethyleneimine, preparing double bonds from the fluorinated polyethyleneimine, and then grafting a targeting peptide E7 to obtain a carrier F-PEI-E7; and (2) entrapping an osteogenesis promoting drug in the carrier F-PEI-E7 to obtain the organic nanoparticles with high osteogenesis performance. The invention provides an innovative strategy: F-PEI is used as a carrier, an osteogenesis promoting drug (beta-GP) is entrapped, and through E7 targeting peptide (EPLQLKM) modification, precise targeting delivery of the drug is realized. The organic nanoparticles are high in stability, the in-vitro osteogenesis promoting effect is equivalent to that of a classical formula for in-vitro osteogenesis induction, and the organic nanoparticles have an excellent in-vivo osteogenesis promoting effect.

The invention belongs to the field of gene engineering. Specifically, the invention relates to a system, a tool and a method for modifying or editing target nucleotide bases, and more specifically, the invention relates to an optimized QBE base editing system and application thereof. The base editing system disclosed by the invention has the advantages of high efficiency of a gene editing tool, high product purity, low Indel frequency, low off-target frequency and abundant gene editing windows, and has important value for expanding application scenes of a base editing technology in the fields of life science basic research, agriculture, biological medicine and the like.

The invention belongs to the technical field of biology, and particularly relates to a cattle TXNIP gene double-target knockout vector as well as a construction method and application thereof. The TXNIP gene double-target knockout vector comprises a lentiviral expression vector, a first sgRNA sequence and a second sgRNA sequence, the first sgRNA sequence is connected to the lentiviral expression vector and targets a bovine TXNIP coding region, the second sgRNA sequence is connected to the lentiviral expression vector and targets the TXNIP coding region, and nucleotide sequences of an upstream primer and a downstream primer corresponding to the first sgRNA sequence are shown as SEQ ID NO.1 and SEQ ID NO.2 respectively. The nucleotide sequences of the upstream primer and the downstream primer corresponding to the second sgRNA sequence are respectively as shown in SEQ ID NO.3 and SEQ ID NO.4. Compared with a single-target knockout vector, the cattle TXNIP gene double-target knockout vector has higher flexibility, the targeting efficiency can be improved, and the off-target effect is reduced.

The invention discloses a multifunctional nano-composite as well as a preparation method and application thereof. The multifunctional nano-composite comprises a DNA carrier, a photothermal agent and an anti-tumor drug, the DNA carrier comprises a long DNA single chain with n repetitive units, n DNA complementary single chains, n aptamer chains with cancer cell targeting capability and n SH-DNA chains connected with photo-thermal agent nanoparticles, wherein n is an integer between 5 and 150; each repetitive unit of the long DNA single chain has 63 basic groups, and the basic groups in each repetitive unit are respectively paired and connected with the DNA complementary single chain, the aptamer chain and the SH-DNA chain. According to the invention, the aptamer with multivalent targeting capability and a large amount of photothermal agents and chemotherapeutic drugs are loaded by utilizing a multi-repetitive unit of the long DNA single-chain carrier, so that a chemotherapy / photothermal / chemical power synergistic treatment system with multivalent aptamer positioning is constructed, and cancer cells can be effectively killed.

The invention relates to the field of gene engineering, and discloses a construction method and application of a GPX4S131A mutant endothelial cell line. According to the invention, the CRISPR / Cas9 technology is utilized to construct a gRNA vector containing targeted GPX4, ssODN is utilized to carry out targeted alanine (A) mutation on serine (S) of the 131 basic group of GPX4, and a stable GPX4S131A mutant endothelial cell ferroptosis model is constructed, so that a specific mechanism of endothelial cell ferroptosis is disclosed, a useful clue is provided for treating cardiovascular diseases caused by endothelial cell ferroptosis, and the application has a wide application prospect. The important application value is realized.

The invention relates to nano delivery, in particular to a photosensitive bionic drug delivery system as well as a preparation method and application thereof. The photosensitive bionic drug delivery system RSCP NPs is mainly composed of a core part and a shell part, wherein the shell is composed of a macrophage membrane RCP co-modified by fat targeting peptide P3 (CKGGRAKDC and a photosensitive supramolecular compound CHD (the compound is composed of DSPE-2-(4-hydroxyphenylazo)-benzoic acid-beta-cyclodextrin); an inner core is pH response type SEP NPs formed by self-assembly of a triblock amphiphilic polymer. The RSCP NPs has higher drug loading capacity, and can respond to multiple signals (inflammatory microenvironment, red light and pH) to regulate the capacity of gradual release of drugs and active targeting of WAT inflammatory areas, so that toxic and side effects caused by off-target effect of the drugs can be effectively reduced, and the effect of improving low-intensity inflammatory microenvironment of obese tissues is achieved.

The invention discloses a PDCD4 inhibitor based on PROTAC modification and application of the PDCD4 inhibitor in preparation of antidepressant drugs, a series of PROTAC modified compounds with a protein degradation function are successfully synthesized by performing PROTAC modification on a small molecule compound ZINC20031600, combining E3 ligase ligands CRBN and VHL and using PEG with different lengths as linkers. On the basis of the prior art, the protein degradation efficiency and selectivity of PROTAC molecules are remarkably improved, the cytotoxicity is reduced, and good clinical application potential and market prospects are achieved.

Provided is a small activating RNA used for promoting an expression level of a target gene. The small activating RNA is composed of a sense oligonucleotide chain and an antisense oligonucleotide chain. A nucleotide of the sense or antisense oligonucleotide chain has 2'-fluoro, 2'-methoxy, and a phosphorothioate modification to the nucleotide backbone.

The invention relates to an antigen and trichosanthin (TCS) co-loaded exosome, a preparation method thereof, an antigen adjuvant integrated vaccine preparation and application. The exosome comprises a cell membrane vesicle, an antigen unit combined on the surface of the cell membrane vesicle, and a trichosanthin unit combined on the surface of the cell membrane vesicle. The exosome can effectively induce generation of antigen-specific antibodies and is beneficial to reduction of off-target effects. The exosome can be used for preparing an antigen adjuvant integrated vaccine preparation, and can be further applied to preparing a medicine for inducing the neutralization of a specific antibody. According to the present invention, the antigen unit of the antibody can be SARS-CoV-2RBD, and the corresponding exosome-like body can effectively induce the maturation of the dendritic cells in vitro, and can effectively induce the generation of the SARS-CoV-2RBD specific antibody in vivo, such that the SARS-CoV-2 infection can be effectively resisted;

The present application relates to an ATP-responsive lipid, a nucleic acid nano-preparation and a preparation method. The structural formula of the ATP-responsive lipid is shown in the following formula (I). The ATP-responsive lipid and the nucleic acid nano-preparation of the present application can be combined with ATP under the condition of high ATP level in cells, increase the fluidity of the membrane, promote the fusion of the lipid nanoparticle and the endosome membrane, destroy the integrity of the endosome membrane, increase the efficiency of lysosome escape, and then improve the efficiency of nucleic acid delivery and expression.

This invention provides the application of a siRNA composition targeting LOC157273 in the preparation of drugs for treating non-alcoholic steatohepatitis (NASH) and liver fibrosis, belonging to the field of biomedical technology. The siRNA of this invention directly corrects the hyperlipidemic hepatic lipid synthesis, insulin resistance, and inflammatory activation caused by abnormal expression of PPP1R3B by specifically silencing the key fibrosis driver gene LOC157273. This fundamentally blocks the activation of hepatic stellate cells and excessive extracellular matrix deposition, achieving precise reversal of the fibrosis process and blocking the occurrence of NASH. The siRNA delivery system is optimized through liver-targeted modification, improving liver enrichment efficiency and reducing off-target effects caused by systemic distribution. It exhibits strong targeting, directly acting on upstream regulatory genes of fibrosis and NASH. Simultaneously, it silences multiple synergistic pathogenic gene sites, enhancing therapeutic efficacy and providing a highly efficient and safe novel intervention for liver fibrosis and NASH.

The invention relates to the technical field of gene editing, in particular to a base editor as well as a construction method and application thereof, which are used for mutation of a specific base at a specific site in DNA (Deoxyribose Nucleic Acid) level. The ABE with higher editing specificity and low RNA / DNA off-target risk, the CBE with high editing precision and low RNA / DNA off-target risk and the ACBE tool with smaller volume are constructed by the method disclosed by the invention. The ABE tool developed in the invention has higher specificity when mediating A.T-G.C mutation, the capability of mediating C.G-T.A mutation of the ABE tool is obviously inhibited and even eliminated, and the off-target effect at RNA and DNA levels is obviously reduced and even eliminated; the CBE tool developed by the invention has higher accuracy when mediating C.G-T.A mutation, and the off-target effect of RNA (Ribonucleic Acid) and DNA (Deoxyribose Nucleic Acid) levels is obviously reduced or even eliminated; the ACBE tool developed in the invention only needs single deaminase, is smaller in volume, obviously reduces and even eliminates off-target effects at RNA and DNA levels, and has higher application value.

The invention provides a self-assembly label-free enzyme-free electrochemical HPV16E7 detection platform as well as a preparation method and application thereof, and belongs to the technical field of biological analysis. According to the invention, a magnetic alpha-Fe2O3 / Fe3O4 (at) Au / Sub / BSA signal transmission medium, a CRISPR / Cas14a system and a double-foot DNA walker are organically combined to prepare a self-assembly label-free enzyme-free electrochemical HPV16E7 detection platform, and a more reliable and accurate means is provided for HPV16E7 detection; the linear detection range of the electrochemical HPV16E7 detection platform is 100 fg.mL <-1 >-10 [mu] g.mL <-1 >, the detection limit is 67.17 fg.mL <-1 >, and the quantification limit is 0.22 pg.mL <-1 >; the electrochemical HPV16E7 detection platform has the advantages of no need of enzyme, no need of marking, no need of pretreatment, fast and convenient detection process, low cost and good practicability.

This invention discloses a transgenic recombinant immune cell that specifically targets tumors. The recombinant immune cell comprises a chimeric polypeptide and a chimeric antigen receptor; the chimeric polypeptide includes a first extracellular region, a first transmembrane region, and a first intracellular region with HLA-G protein binding activity, wherein the N-terminus of the first transmembrane region is connected to the C-terminus of the first extracellular region, and the N-terminus of the first intracellular region is connected to the C-terminus of the first transmembrane region; the first transmembrane region includes the transmembrane region of the Notch receptor protein from Xenopus laevis; the antigen-chimeric receptor does not have HLA-G protein binding activity. This recombinant immune cell significantly improves the precise recognition of tumor cells by immune cells, reduces off-target effects of cell therapy, and accurately and effectively distinguishes tumor cells from normal cells, providing an effective target and targeting method for the treatment of broad-spectrum tumors (especially solid tumors).

The invention provides application of ANKRD13A in preparation of drugs and diagnostic reagents for intestinal cancer, relates to the technical field of biomedicine, and is technically characterized by providing application of ANKRD13A as a therapeutic target in preparation of drugs for preventing and / or treating intestinal cancer. Close association between ANKRD13A and occurrence and development of the intestinal cancer is defined through a series of verification experiments. Clinical sample analysis shows that transcript abundance and protein expression level of ANKRD13A in a colon cancer tissue are remarkably lower than those in a normal colon tissue; the cell level verification proves that the mRNA and protein expression of ANKRD13A in the colon cancer cell line is obviously reduced compared with that of a normal colon epithelial cell line; animal experiments show that an Ankrd13a gene-deficient mouse is induced by AOM / DSS, the occurrence rate of colorectal tumors is higher, the number of the tumors is larger, and tumor tissues have medium-high-grade canceration characteristics along with abnormal expression of beta-Catenin, nuclear ectopic, increase of Ki67 positive cells and other malignant pathological expressions. The experimental results jointly prove that expression deletion or down-regulation of ANKRD13A is closely related to occurrence risk increase of intestinal cancer.

The present invention is directed to provide novel RNA molecules. The RNA molecules are for RNA interference to target a mutant allele with a point mutation, in which (1) the molecule has a nucleotide sequence complementary to a nucleotide sequence of a coding region of the mutant allele; and (2) when counted from the base at the 5'-end in the nucleotide sequence complementary to the nucleotide sequence of the mutant allele, (2-1) a base at position 5 or 6 is mismatched with a base in the mutant allele; (2-2) a base at position 10 or 11 is at the position of the point mutation and is identical to the base at the position of the point mutation in the mutant allele; (2-3) the group at the 2'-position of the pentose in the ribonucleotide at position 8 is modified with OCH3, halogen, or LNA; and (2-4) the group at the 2'-position of the pentose in the ribonucleotide at position 7 is not modified with any of OCH3, halogen, and LNA. In this RNA molecule, one or more ribonucleotides may be each replaced by, e.g., a deoxyribonucleotide. The molecule may form a double-stranded RNA with a complementary strand.

The invention relates to a plasmid-loaded targeted liposome compound, and a preparation method and application thereof. The liposome compound comprises liposome and plasmid pPEA entrapped by the liposome, and the plasmid pPEA comprises an expression vector pGAVPO containing a blue light induced transcription factor GAVPO protein and pU5-PEA of a light-operated gene expression system capable of expressing PEA. The targeting biological material for modifying the liposome is fused on the liposome to form a liposome compound pPEA (at) GPH-L entrapping the pseudomonas aeruginosa exotoxin light-operated gene and the liposome targeting material. The liposome disclosed by the invention can target tumor cells, has great lethality on the tumor cells, is safe and effective, and has important significance in preparation of anti-tumor medicines, especially medicines for treating breast cancer.

The invention relates to the technical field of gene editing, in particular to a base editor as well as a construction method and application thereof, which are used for mutation of a specific base at a specific site in DNA (Deoxyribose Nucleic Acid) level. The ABE with higher editing specificity and low RNA / DNA off-target risk, the CBE with high editing precision and low RNA / DNA off-target risk and the ACBE tool with smaller volume are constructed by the method disclosed by the invention. The ABE tool developed in the invention has higher specificity when mediating A.T-G.C mutation, the capability of mediating C.G-T.A mutation of the ABE tool is obviously inhibited and even eliminated, and the off-target effect at RNA and DNA levels is obviously reduced and even eliminated; the CBE tool developed by the invention has higher accuracy when mediating C.G-T.A mutation, and the off-target effect of RNA (Ribonucleic Acid) and DNA (Deoxyribose Nucleic Acid) levels is obviously reduced or even eliminated; the ACBE tool developed in the invention only needs single deaminase, is smaller in volume, obviously reduces and even eliminates off-target effects at RNA and DNA levels, and has higher application value.

The invention belongs to the technical field of gene engineering and biology, and relates to screening, construction and application of a modified AAV-8 serotype for gene targeting and expression. The invention discloses a modified type of an AAV-8 serotype. An amino acid sequence as shown in SEQ ID NO.5 is inserted between the 590th amino acid and the 591th amino acid of a capsid protein sequence of the AAV-8 serotype as shown in SEQ ID NO.2. The invention further discloses a preparation method of the modified type of the AAV-8 serotype. The invention also includes corresponding nucleic acids and capsids and uses thereof. The modified serotype disclosed by the invention is stronger in fluorescence effect after infection and can be obviously and intuitively observed, an exogenous gene can be widely and efficiently expressed in a plurality of areas such as a targeted tissue retina outer nuclear layer, a light sensation cell layer and a retinal pigment epithelial cell layer, and under the condition that the same virus amount is used, the expression efficiency is greatly improved. And compared with a wild serotype, the serotype has better and wider targeted tissue expression and less non-targeted tissue expression.

The present invention discloses a drug delivery nanosystem and method based on DNA origami and doxorubicin prodrug, belonging to the field of biomedical technology. In view of the abnormally high expression of NQO1 enzyme in many tumor cells, the present application synthesizes a target-activated anti-tumor prodrug based on doxorubicin as a model, and its pharmacological action can be activated by the NQO1 enzyme; at the same time, a rectangular DNA origami with good biocompatibility is used as a nanocarrier to efficiently carry the prodrug, thereby constructing an activatable precision drug delivery nanosystem. This prodrug-embedded rectangular DNA origami is conducive to reducing the toxic and side effects of anticancer drugs on normal human liver cells (L02 cells), and has a significant inhibitory effect on human breast cancer cells (MCF-7 cells) with high expression of NQO1 enzyme. This therapeutic nanosystem is expected to reduce the systemic toxicity of tumor drugs due to non-specificity, improve patients' compliance and quality of life, and provide an effective strategy for precise clinical intervention of tumors.

The invention belongs to the technical field of gene engineering and biology, and relates to screening, construction and application of a modified AAV-8 serotype for gene targeting and expression. The invention relates to a polypeptide, in particular to a modified AAV-8 serotype for gene targeting and expression, which is characterized in that a polypeptide consisting of 10 amino acids is inserted between the 590th amino acid and the 591th amino acid of the sequence of AAV-8 serotype capsid protein as shown in SEQ ID NO.2, the polypeptide composed of 10 amino acids contains the polypeptide with the amino acid sequence as shown in SEQ ID NO. 5. After the modified serotype is infected, the fluorescent effect is stronger, obvious and visual observation can be realized, an exogenous gene can efficiently and widely express a target tissue retina outer nuclear layer and a light sensitive cell layer in vivo, and wider target tissue expression and less non-target tissue expression are realized.

The invention relates to the technical field of medicines, in particular to NUDT5 and SEPT8 serving as novel prostate cancer treatment targets and application of the NUDT5 and the SEPT8 in prostate cancer treatment. The invention provides a novel targeted therapy scheme for prostatic cancer, the prostatic cancer cells are specifically killed in vivo by inhibiting prostatic cancer cell proliferation by simultaneously targeting NUDT5 and SEPT8 proteins with relatively high expression quantity in high-risk prostatic cancer, and a new thought is provided for clinical treatment of prostatic cancer.

The invention belongs to the technical field of gene engineering and biology, and relates to screening, construction and application of a modified AAV-8 serotype for gene targeting and expression. According to the modified adeno-associated virus AAV-8 serotype for retina outer nuclear layer targeting and expression, 10 amino acid sequences shown as SEQ ID NO.3 are inserted between the 590th amino acid and the 591th amino acid of AAV-8 serotype capsid protein, the amino acids at the first, second and tenth sites are protected amino acids, and the amino acids at the first, second and tenth sites are protected amino acids. The third to ninth amino acids are amino acid sequences obtained by screening. The modified serotype provided by the invention has a stronger fluorescence effect after infection, can obviously and intuitively observe that an exogenous gene can be widely and efficiently expressed in a specific targeted tissue retina outer nuclear layer area, and has better and wider targeted tissue expression than a wild serotype under the condition of using the same virus amount.

The present invention relates to the field of nucleic acid editing, in particular to the technical field of regularly clustered interspaced short palindromic repeats (CRISPR). Specifically, the present invention relates to Cas effector proteins, fusion proteins comprising such proteins, and nucleic acid molecules encoding them. The present invention also relates to complexes and compositions for nucleic acid editing (e.g., gene or genome editing), comprising proteins or fusion proteins of the present invention, or nucleic acid molecules encoding them. The present invention also relates to methods for nucleic acid editing (e.g., gene or genome editing), which use proteins or fusion proteins comprising the present invention.

The invention provides a heart failure treatment device based on an ultrasonic super lens, which comprises a transmitting module and a phase modulation module, the transmitting module comprises a transducer, the phase modulation module comprises the ultrasonic super lens, the transducer and the ultrasonic super lens are tightly attached, the central points of the transducer and the ultrasonic super lens are aligned, and the transducer can transmit a plane wave signal to be modulated. The ultrasonic super lens modulates a plane wave sound field to form a non-diffractive symmetrical Airy sound beam, and generates a focused ultrasonic field at a sub-wavelength focus. Ultrasonic waves are accurately focused on the left ventricle, so that accidental injuries to surrounding sensitive tissues are reduced to the maximum extent, the off-target effect is reduced, and accurate treatment can be achieved without other invasive or complex means. The traditional Chinese medicine composition can effectively reduce myocardial fibrosis and improve cardiac functions, and does not need invasive surgery. In addition, the risk of arrhythmia can be reduced, and long-term and safe heart treatment is achieved.

The invention relates to an application of a modified nucleotide monomer in nucleic acid interference (RNAi) drug molecules. The RNAi molecule (or oligomeric nucleic acid) carrying the nucleotide monomer disclosed by the invention is a double-stranded siRNA molecule. According to the present invention, the nucleotide monomer is introduced into the antisense strand seed region of siRNA through the specific 2 ', 5'-phosphate linking, such that the potential off-target effect of the antisense strand seed region can be significantly reduced, such that the siRNA can highly selectively silence the target gene mRNA, the siRNA treatment safety window is improved, and the related diseases caused by the target gene mRNA can be effectively treated.

The invention discloses a sinomenine derivative and a synthesis method and application thereof, and belongs to the technical field of biology and medicines.The sinomenine derivative is synthesized through the steps that sinomenine and acryloyl chloride are subjected to a room-temperature reaction under the catalysis of triethylamine, then a product in reaction liquid is separated and purified, and an intermediate is obtained; the intermediate and substituted aniline are subjected to a room temperature reaction for 11-13 h under the catalysis of organic alkali, then a product in reaction liquid is separated and purified, and the sinomenine derivative is obtained and can be used as an antifungal agent or applied to preparation of antifungal drugs, and the sinomenine derivative and fluconazole are combined to be used as an antifungal drug synergist. The sinomenine derivative not only provides a new candidate drug for fungal treatment, but also improves the antifungal effect of the existing drug.

The invention provides a polypeptide hydrogel. The polypeptide hydrogel is an AG73-GelMA / AlgMA hydrogel which is prepared by crosslinking an AG73 polypeptide and a GelMA / AlgMA hydrogel. The invention further provides a preparation method of the polypeptide hydrogel. The hydrogels are used to produce pancreatic progenitor cell (PP) organoid and to improve the SC-beta cell differentiation regimen. AG73 can simulate the interaction between cells and a matrix, activate an AKT signal channel, promote the formation of PDX1 + / NKX6.1 + PP organs, and effectively regulate subsequent epithelial-mesenchymal transition (EMT) in an endocrine lineage; the GelMA / AlgMA hydrogel provides optimal mechanical stress and space structure for differentiation of PP (propene polymer) organs by simulating the physiological stiffness of pancreas. The SDC4-ITGAV compound plays a key role in the early stage of pancreatic development, and promotes the formation of SOX9 + / PDX1 + dual-energy PPs. The SC-beta cell derived from the AG73-GelMA / AlgMA hydrogel disclosed by the invention shows that the insulin secretion function is enhanced in vivo, and meanwhile, the hyperglycemia state of a diabetic mouse is more quickly reversed. The invention provides a cost-effective, stable and efficient alternative scheme for the culture of in vitro SC-beta cells by reducing the uncertainty related to the traditional culture method.

The present invention belongs to the field of medicine and biology, and specifically relates to a dual-target CAR vector combined with EpCAM and MSLN single-chain antibodies, a construction method thereof, and an application in ovarian cancer. The dual-target CAR vector combines EpCAM single-chain antibodies and MSLN single-chain antibodies, and the specific composition structure of CAR is CD8leader-EpCAM scFv-(G4S)5-MSLN scFv-CD8α-CD28-CD137-CD3ζ. CAR-T cells (Chimeric Antigen Receptor T-Cell) containing the therapeutic vector are obtained by viral infection, and by expressing the CAR structure, ovarian cancer cells that highly express EpCAM and MSLN tumor-associated antigens can be targeted for recognition and killing.