17 results about "Ribozyme" patented technology

The present invention relates to a method for producing a circular gene of interest. A nucleic acid molecule comprising in the 5' to 3' direction a first bridging sequence and a gene of interest is provided. The first bridging sequence comprises a sequence corresponding to the 3' part of a ribozyme. A modified ribozyme comprising a truncated ribozyme which does not comprise the 3' part of the corresponding wild type ribozyme is also provided. The nucleic acid molecule and the modified ribozyme are combined under conditions suitable for circularization to occur. The invention also provides a modified ribozyme, a nucleic acid molecule and a kit.

The application provides a plasmid containing modified satellite RNA and a construction method and application, and belongs to the field of plant virology and molecular biology. Each copy number of the modified satellite RNA is composed of a modified satRNA TA-Tb fragment and an HDV ribozyme fragment, and the copy number of the modified satellite RNA in the plasmid is 10. The nucleotide sequence of the modified satellite RNA is shown in SEQ No. 3. After the plasmid of the application is inoculated by agrobacterium injection, the symptoms caused by cucumber mosaic virus on different strains and the same strain (CMV Fny ) on different hosts can be significantly reduced. An effective control measure for field control of cucumber mosaic virus is provided.

The technology described herein is directed to Aptamer and Ribozyme Equilibrium Shifting (ARES) regions, including ON-switches and OFF-switches, which can be harnessed to regulate the stability of RNA molecules. Also described herein are compositions comprising such RNA molecules and methods of using them to regulate translation of cargo polypeptides.

Provided are methods and compositions for screening / identifying / characterizing nucleic acid delivery vehicles (e.g., liquid nanoparticle (LNP) formulations). Provided is a library of nucleic acid delivery vehicles with different chemical compositions, where the members of the library are barcoded by virtue of including a barcoded RNA molecule (in some cases a barcoded ligation-ready RNA molecule). The barcoded RNA molecule includes a first ribozyme, a second ribozyme, and a barcode sequence between the first and second ribozymes. Once a barcoded RNA molecule is successfully delivered into a cell, the first and second ribozymes cleave the barcoded RNA molecule to produce a 5'-OH end and a 2',3'-cyclic phosphate end, which ends can be ligated to one another by an intracellular RNA ligase, producing a circular RNA (cRNA) that includes the barcode sequence. Detecting barcode sequences from cRNAs therefore identifies the chemical composition of successful delivery vehicles.

The application discloses a kind of photoactivation manganese ion bridging type deoxyribozyme probe preparation and its application in Cistanche inhibiting acetylcholinesterase activity.The probe is pSD@MnO2 Composite structure, which is composed of manganese dioxide nanosheet and functional nucleic acid assembly adsorbed on its surface, the assembly includes S chain labeled with fluorescence / quenching group and containing deoxyribozyme cleavage site, and pD chain partially hybridized with S chain and containing photocleavage site.Its preparation includes nucleic acid probe assembly, MnO2 Nanosheet complexing and purification steps.When applied, AChE catalytic product decomposes MnO2, releases Mn 2+ And nucleic acid chain; after centrifugation, pD chain is cut by ultraviolet light activation trigger, and released DNAzyme is cut under the assistance of Mn 2+ It produces amplified fluorescence signal; by comparing signal change before and after adding inhibitor, inhibition activity can be quantitatively evaluated.The application combines photocontrol activation with enzyme cascade amplification, realizes high sensitivity and operation synchronism of detection process, and is especially suitable for high-throughput rapid screening of AChE inhibitor in Cistanche and other natural products.

This invention discloses a bisphosphonate-nucleic acid nanoparticle, its preparation method, and its applications. The preparation method involves adding divalent metal ions to a rolling circle amplification (RoBA) reaction solution, followed by the addition of bisphosphonate, to perform a RoBA reaction to obtain the bisphosphonate-nucleic acid nanoparticles. The RoBA reaction solution contains a circular DNA template. The preparation method of this invention has the advantages of high speed, precise synthesis, and simplicity, realizing multiple reactions—RoBA, ion coordination, crystallization, and co-assembly of DNA and bisphosphonate—in the same reaction system, greatly simplifying the reaction process and improving reaction efficiency. The obtained bisphosphonate-nucleic acid nanoparticles exhibit dual-pathway therapeutic effects and excellent ribozyme stability, showing promising application prospects.

This invention belongs to the medical field and provides a calcium carbonate manganese nanoprobe for in situ imaging detection of vascular epidermal growth factor, its preparation method, and its application. In this strategy, a DNA strand (P1 and P2) containing a protein recognition antibody, a protein recognition sequence, and a deoxyribonuclease, along with a molecular beacon DNA strand (H1), is stirred in a DMEM cell culture medium containing calcium chloride and manganese chloride to prepare the calcium carbonate manganese nanoprobe. In acidic tumor cells, this nanoprobe releases P1, P2, H1, and manganese ions. P1 and P2 can simultaneously recognize proteins, forming a complete deoxyribonuclease center, which catalyzes the cleavage of H1 under the action of the cofactor manganese ions, generating a fluorescent signal. This strategy achieves specific in situ dynamic imaging of low-abundance proteins in vivo and has great potential in biomedical research.

This invention discloses a deoxyribozyme probe for recognizing strontium ions and its applications, belonging to the field of deoxyribozyme probe technology. The nucleotide sequence of the deoxyribozyme probe provided by this invention is any one of the sequences shown in SEQ ID NO:1 to SEQ ID NO:9. Specifically, it is a cis-structured deoxyribozyme probe composed of a deoxyribozyme and a substrate chain with a fluorescent group and a quenching group. The substrate chain sequence is shown in SEQ ID NO:10, and the deoxyribozyme sequence is any one of the sequences shown in SEQ ID NO:17 to SEQ ID NO:25. The deoxyribozyme probe provided by this invention can efficiently and specifically recognize strontium ions, thereby achieving accurate detection of the target ion, with a strontium ion response concentration of 2 mM or higher. This probe has good application prospects in the fields of biosensing and ion detection.

A flexible method for constructing a vector for preparing circular RNA without being limited by substrate sequence is provided. The method comprises the following steps: determining a target site sequence NNUNNNN to undergo cyclization, and dividing said sequence into E1: NNU and E2: NNNN; using the E1 and E2 sequences as references to design IGS sequences to form complementary pairings with E1 and E2, respectively; adding 5' and 3' homologous arms, as well as IRES and CDS and other elements, respectively. In the provided method, circular RNA can be prepared for any target sequence without residual sequences, and the described method has high cyclization efficiency. Also provided is a FlexCirc ring-forming system designed on the basis of an Azoarcus group I intron ribozyme, which can form circular RNA. The ring-forming substrate sequence has the characteristics of flexible design, and the ring-forming efficiency can achieve a high ring-forming ratio.

This invention provides a novel high-throughput single-cell multi-omics analysis method, comprising the following steps: fixation, lysis, nuclear permeation, and enzyme digestion of a population of cells; adding an A-tail and adjacent ligation to the DNA ends; and performing a transposition reaction. The pretreated cells are loaded onto a microfluidic platform, where single or multiple cells are co-encapsulated in droplets with beads containing capture adapters and a first molecular tag. The DNA and RNA of the same cell are simultaneously labeled using the capture adapters. The cells are released from the droplets and dispensed into multi-well plates. After decrosslinking, lysis, and DNA purification, primers with a second molecular tag are added to simultaneously amplify gDNA and cDNA, forming a unique composite tag. Library construction and sequencing are then performed. This invention improves cell throughput and data quality in a single experiment through a combined indexing approach, addressing the problems of low throughput, high cost, and significant ATAC signal background noise in existing single-cell three-omics technologies.

The disclosure features compositions, systems, and methods for preparation and use of efficient RNA nuclear export of ribozyme-assisted circular RNA molecules (racRNAs). In embodiments, the methods involve characterizing a cell or tissue using racRNAs.

The present invention is directed to a composition for a target sequence detection in double- stranded DNA, wherein the composition comprises i) a first oligonucleotide and a second oligonucleotide for a multi-component deoxyribozyme (MNAzyme), wherein said first and second oligonucleotides are capable of forming substrate binding arms, a catalytic core, and target-binding arms of said MNAzyme, said target-binding arms being complementary to said target sequence in double-stranded DNA, ii) a peptide nucleic acid (PNA) comprising a sequence identical to said target sequence, wherein said PNA is capable of invading the double-stranded DNA at the site of said target sequence and opening the double-stranded DNA by binding to the sequence complementary to said target sequence in said double- stranded DNA leaving said target sequence single-stranded and thus free for said target- binding arms of said MNAzyme to bind, and iii) a substrate oligonucleotide comprising a sequence complementary to the sequence of said substrate binding arms, wherein said substrate binding arms of said MNAzyme engage said substrate oligonucleotide so that said catalytic core of said MNAzyme is capable of modifying said substrate oligonucleotide when said target-binding arms of said MNAzyme bind or are bound to said target sequence in the presence of a cofactor. The present invention is also directed to a kit comprising said composition and to methods of using said composition.

The application discloses a ctPistol ribozyme and application thereof in gene silencing, and belongs to the technical field of biology.The disclosed ctPistol ribozyme can realize effective control of cell phenotype by silencing the expression of key genes.In addition, when applied in a mouse cancer model, the ctPistol realizes a powerful and persistent anti-tumor effect by targeting oncogenic driver genes.As a powerful gene silencing tool and a promising RNA-based therapeutic candidate drug, the ctPistol ribozyme shows an effective way of developing diversified ribozyme-mediated gene control technology.

The application discloses a specific RNA-m 1 The application discloses a chain deoxyribozyme probe for specifically recognizing RNA-m 1 The application discloses a chain deoxyribozyme probe for specifically recognizing RNA-m 1 The application discloses a chain deoxyribozyme probe for specifically recognizing RNA-m

The application provides a preparation method of an engineered DEAR nucleic acid manipulation system, and the preparation method comprises at least one selected from the following: lengthening a substrate recognition region in a RNA molecule of an original DEAR nucleic acid manipulation system to have a length of 7-14 nucleotides; adding a recruitment sequence to a 3' end of a RNA molecule of the original DEAR nucleic acid manipulation system; respectively arranging a first dimerization motif and a second dimerization motif in a III domain in a first RNA molecule and a second RNA molecule from the original DEAR nucleic acid manipulation system, so that the first RNA molecule and the second RNA molecule form a heterodimer, and the like. The preparation method of the engineered DEAR nucleic acid manipulation system provided by the application further improves the specificity and cleavage activity of the DEAR nucleic acid manipulation system based on an RNA ribozyme.

The present application relates to the field of tumor immunotherapy, discloses a kind of ribozyme for targeted inhibition of PD-L1 gene expression and its preparation method and application, specifically relates to a kind of engineered Pistol ribozyme and its use in targeted inhibition of PD-L1 (CD274) gene expression.The ribozyme is RNA molecule connected by ribonucleotide monomer, contains the substrate recognition region of the conserved structural framework maintaining catalytic activity and PD-L1 transcript complement;Conserved skeleton covers P1, P2, P3 region and false node area, and contains catalytic related conserved base site, substrate recognition region can be designed as required, can recognize and cut the targeted site containing GU dinucleotide in PD-L1 transcript, reduce the PD-L1 expression of tumor cell in post-transcriptional level.The PD-L1 transcript includes mature mRNA, precursor mRNA, intron retention transcript and variable splicing transcript.Different from existing immune checkpoint blockade therapy, the present application can sustainably inhibit endogenous PD-L1 in tumor cell, relieve immune suppression, enhance anti-tumor immune response, and also provide the preparation method of the ribozyme and its pharmaceutical composition, and application prospect is good.