8 results about "Rna hybridization" patented technology

The invention relates to the technical field of biological medicine, in particular to a molecular marker of alveolar rhabdomyosarcoma and application of the molecular marker, the molecular marker is a circPAX3R-loop site, and a DNA-RNA hybridization region of the molecular marker is located on chr2: 2230 94913-2230 95853 of a human genome. According to the application disclosed by the invention, a large number of R-loop structures exist in an alveolar rhabdomyosarcoma cell through DRIP-seq, a plurality of circRNAs exist in a cell nucleus of the alveolar rhabdomyosarcoma through circRNA sequencing, the existence of circPAX3R-loop is proved through qPCR after a DRIP product is subjected to RNase R and RNase H treatment, and the result shows that circPAX3R-loop exists in the cell nucleus of the alveolar rhabdomyosarcoma in the cell nucleus of the alveolar rhabdomyosarcoma in the cell nucleus of the alveolar rhabdomyosarcoma. The important clinical application value is realized on developing a reagent or a kit for diagnosing the alveolar rhabdomyosarcoma, a medicine for preventing and / or treating the alveolar rhabdomyosarcoma or a gene treatment strategy aiming at the target spot.

The present application relates to the technical field of biological medicine, and in particular to a molecular marker of alveolar rhabdomyosarcoma and application thereof, wherein the molecular marker is circFOXO1R-loop site, and a DNA-RNA hybridization region thereof is located at chr13:41133915-41134263 of a human genome. In the present application, the circFOXO1R-loop site is verified by CUT&Tag-qPCR and DRIP-qPCR, and it is found that the circFOXO1R-loop can promote the combination of FOXO1 gene and RNAPol II and DNA damage, and it has important clinical application value for developing a reagent or kit for diagnosing alveolar rhabdomyosarcoma, a drug or a gene therapy strategy for preventing and / or treating alveolar rhabdomyosarcoma aiming at the target.

This invention relates to an in situ multi-throughput glycosylated RNA imaging method, comprising the following steps: constructing different proxHCR systems for different glycoRNAs, thereby performing multi-throughput in situ imaging of multiple glycoRNAs with the same sialic acid composition but different RNA sequences; the proxHCR system includes a sialic acid recognition module, an RNA hybridization module, and a fluorescently labeled hairpin probe (AR) acting as an amplification reporter. The sialic acid recognition module and the proxHCR module are used for dual recognition of sialic acid and RNA units of glycoRNAs on the cell membrane surface. A complete activation foothold switch is formed only when the sialic acid and RNA units of the glycoRNAs on the cell membrane surface are spatially adjacent, triggering HCR of the fluorescently labeled hairpin probe (AR), thereby visualizing the glycoRNAs on the cell surface through amplified fluorescence signals. This invention solves the problem of the lack of prior art for simultaneous multi-throughput analysis of multiple different glycosylated RNAs.

The invention discloses a stimuli-responsive hepatic cell targeted light-controlled nano-carrier UACPG and a preparation method and application thereof, the preparation method comprises the following steps: extending the 3'end of sgRNA to synthesize sgRNAL, the extending part is an ASO base complementary pairing sequence, and then preparing Cas9 / sgRNA / ASO through DNA-RNA hybridization; the preparation method comprises the following steps: preparing Nd < 3 + >-sensitized core-shell UCNPs nanoparticles which take NaYF4: Tm / Yb / Ca as a core and NaYF4: Nd / Yb as a shell; dBCO group modification is carried out on the UAC nano particles, Cas9 / sgRNA / ASO is efficiently coupled through click chemistry, and UAC nano particles are synthesized; finally, the surfaces of the UAC nano particles are coated with galactosylated polyacetamide (PVAm-Gal), and the UACPG nano particles are obtained. Safe, efficient and high-targeting delivery of Cas9 / sgRNA / ASO in vivo and in vitro is realized; and the traceless release of ASO and Cas9 / sgRNA is realized under the irradiation of NIR, and the purposes of blocking virus replication, supplementing in a cccDNA pool and removing the existing cccDNA are achieved by targeting HBV pgRNA and HBV cccDNA respectively.

The invention belongs to the technical field of biology, and discloses a capturing method of ready-to-use RNA (Ribonucleic Acid) in-situ interaction protein. The method comprises the following steps: S1, adding a biotin-labeled FISH probe into a cell suspension; s2, then adding streptavidin to be labeled with horse radish peroxidase (Streptavidin-HRP), and carrying out enzyme deactivation on the horse radish peroxidase; s3, washing off non-specific binding; and S4, adding biotin phenol and a hydrogen peroxide substrate to trigger a biotinylation labeling reaction. According to the method, a ready-to-use RNA in-situ proximity biotinylation labeling technology without genetic manipulation is established through fusion RNA-FISH in-situ hybridization and proximity labeling technologies, proximity labeling enzyme can be directly guided to interested RNA through an RNA hybridization probe in a fixed cell through the technology, then a proximity biotinylation labeling reaction is carried out, and the RNA in-situ proximity biotinylation labeling technology is established. The method comprises the following steps of: adding a biotin label to interacting protein adjacent to RNA (Ribonucleic Acid), so that the protein can be enriched by streptavidin-beads for mass spectrum identification.

The invention provides a method for measuring the positive rate of cells expressing exogenous therapeutic genes by combining RNA (Ribonucleic Acid) hybridization and flow cytometry. The detection method comprises the following steps: designing a specific in-situ hybridization probe aiming at mRNA (Messenger Ribonucleic Acid) expressed by a target gene, carrying out RNA probe in-situ hybridization staining in the cells, and distinguishing specific cell subgroups by combining the single cell resolution of flow cytometry. A feasible detection method is provided for highly personalized and multivalent TCR and other cell treatment samples which do not have specific antibodies or cannot detect the positive rate by using a unique antibody, and the design space for designing the specific target RNA probe is larger and the difficulty is lower than that for designing the specific antibody. Comprising the following steps: fixing cell morphology and RNA, permeabilizing / breaking a cell membrane, fixing / perforating a cell nucleus, digesting intracellular and intracellular free single-stranded DNA, carrying out RNA probe hybridization and signal amplification, and carrying out flow detection.

The invention relates to the technical field of biological medicine, in particular to a molecular marker for alveolar rhabdomyosarcoma and application of the molecular marker, the molecular marker is a circFOXO1R-loop site, and a DNA-RNA hybridization region of the molecular marker is located on chr13: 41133915-41134263 of a human genome. According to the invention, the method comprises the following steps: CUTamp; the circFOXO1R-loop site is verified through Tag-qPCR and DRIP-qPCR, it is found that the circFOXO1R-loop can promote combination of the FOXO1 gene and RNAPol II and DNA damage, and the circFOXO1R-loop has important clinical application value for developing a reagent or a kit for diagnosing the alveolar rhabdomyosarcoma, a medicine for preventing and / or treating the alveolar rhabdomyosarcoma or a gene treatment strategy aiming at the target site.

The present invention belongs to the technical field of nanocarriers, and relates to a nanocarrier based on a DNA-RNA hybrid molecule, its preparation method and application. The nanocarrier includes a DNA-RNA hybrid molecule, an AS1411 aptamer and a target RNA. The DNA-RNA hybrid molecule is a connecting bridge between the AS1411 aptamer and the target RNA. By using DNA-RNA hybridization technology, a DNA-RNA hybrid nanocarrier delivery system carrying siRNA / saRNA is constructed. This delivery system has extremely low cytotoxicity and excellent targeting compared with traditional transfection reagents, can successfully target and deliver the target small RNA into tumor cells, and up-regulate or down-regulate the expression of its target gene, inducing apoptosis of tumor cells, thereby playing an anti-tumor role. The construction of this delivery system provides a new idea for the development of small RNA delivery systems.