353 results about "Circular RNA" patented technology

The present invention relates to a single-chain circular RNA having a sustained or slow-releasing RNA interference effect, characterized in that the single-chain circular RNA comprises a sense strand sequence, an antisense strand sequence complementary to the sense strand sequence, identical or different two loop sequences between the sense strand and the antisense strand, connecting both strands, wherein the sense strand and the antisense strand are paired to form a stem.

Currently, the circularization of small RNAs is broadly regarded as an obstacle in ligation-related assays and explicitly avoided while short lengths of linear RNA targets is broadly recognized as a factor limiting use of conventional primers in PCR-related assays. In contrast, the disclosed invention capitalizes on circularization of small RNA targets or their conjugates with oligonucleotide adapters. The circular RNA templates provide amplification of the target sequences via synthesis of multimer nucleic acids that can be either labeled for direct detection or subjected to PCR amplification and detection. Structure of small circular RNAs and corresponding multimeric nucleic acids provide certain advantages over current methods including flexibility in design of conventional RT and PCR primers as well as use of 5′-overlapping dimer-primers for efficient and sequence-specific amplification of short target sequences. Our invention also reduces number of steps and reagents while increasing sensitivity and accuracy of detection of small RNAs with both 2′OH and 2′-OMe at their 3′ ends. Our invention increase sensitivity and specificity of detection of microRNAs and other small RNAs with both 2′OH and 2′-OMe at their 3′ ends while allowing us to distinguish these two forms from each other.

The present invention relates to inhibition of microRNAs (miRNA), other types of RNA and RNA-binding proteins. In particular the present invention relates to nucleic acid sequences that are circular and allow inhibition of the interactions between microRNAs and their RNA target and being resistant to degradation by exonucleases. Also, the present invention relates to nucleic acid sequence that are circular and acts as an RNA blocker or protein decoy.

The invention discloses an establishing method of a circular RNA high-throughput sequencing library and a kit thereof. The establishing method sequentially comprises the following steps: (S1) extracting total RNA from a sample; (S2) removing DNA in the sample; (S3) detecting and evaluating the quality of the total RNA in the sample, and determining that the RNA quality meets a requirement; (S4) removing rRNA; (S5) removing linear RNA; (S6) constructing the circular RNA library for high-throughput sequencing. The establishing method of the circular RNA high-throughput sequencing library, provided by the invention, is high in efficiency, stable, low in residual ratio of the rRNA, high in circular RNA detection efficiency, good in data reproducibility, and high in sequencing result verifying success rate, and is especially applicable to an FFPE tissue and other samples with poor RNA quality.

The invention discloses a circular RNA-MET gene in liver cancer as well as an expression method and a fluorescent quantitative PCR (polymerase chain reaction) detection method of the circular RNA-MET gene. The cDNA nucleotide sequence of the circular RNA-MET gene in the liver cancer is shown by SEQ ID NO:1. By designing a specific primer capable of amplifying the circular RNA, the circular RNA of the MET gene is amplified out by PCR, and an accurate cyclization site of circular RNA-MET is measured by a method for sequencing a PCR product namely cloned DNA; and the MET gene is determined to objectively contain the circular RNA consisting of 1214 nucleotides. By using the fluorescent quantitative PCR detection method of the circular RNA-MET gene in the liver cancer, the existence and expressive differences of the circular RNA-MET in samples of liver cancer cells and liver cancer clinical tissues can be specifically detected. The invention can provide an ideal gene detection method for early clinical diagnosis of liver cancer.

The invention discloses a method for screening specific circular RNA (ribonucleic acid) of triple-negative breast cancer. The method includes comparing circular gene expression of triple-negative breast cancer plasma samples and normal comparison groups by the aid of 70 parts of specific circular RNA of breast cancer; screening the specific circular RNA to obtain the circular RNA with functions of specifically obviously expressing the triple-negative breast cancer. The specific circular RNA is used as primers. The method has the advantage that results obtained by the aid of the method have important significance on triple-negative breast cancer diagnosis and treatment.

The invention discloses an application of circular RNA WHSC1 in preparation of clinical diagnosis markers in colorectal cancer tumorigenesis and hepatic metastases diseases. The invention further discloses an application of the circular RNA WHSC1 serving as a therapeutic target of the colorectal cancer drug. The invention further discloses a preparation for treating colorectal cancer. The preparation comprises a sequence for specifically interfering the circular RNA WHSC1, wherein the sequence is used for inhibiting the level of the circular RNA WHSC1. Research results show clinical significances of the circ-WHSC1 in diagnosis of colorectal cancer and metastasis and an application in preparation of drugs for treating the colorectal cancer metastasis.

The invention relates to intron-source circular RNA molecules and an application of cyclization key nucleotide sequences of intron-source circular RNA molecules. The invention specifically discloses sequences of circular RNA novel molecules and genome mapping and the cyclization key nucleotide sequences of the circular RNA novel molecules and further discloses a set of plasmids which contain the cyclization key nucleotide sequences for generating the circular RNA molecules comprising any nucleotide sequences. The invention discloses the intron-source circular RNA molecules and the application of cyclization key nucleotide sequences of intron-source circular RNA molecules for the first time, thereby providing a brand-new concept and method for generating the circular RNA molecules comprising any nucleotide sequences.

A method of amplifying DNA from RNA in a sample by using circular RNA is provided.

The invention establishes a circRNA expression profile for a patient with craniosynostosis, provides, provides serum circRNA marker hsa_circ_0001788 for the patient with craniosynostosis, discloses diagnostic value of hsa_circ_0001788 of serum for craniosynostosis, applies the marker herein to prepare a diagnostic kit for the patient with craniosynostosis, and provides supporting for clinical early discovery and treatment of the patient with craniosynostosis. Serum circRNA has good stability, degradation difficulty and the other advantage, has high value in auxiliary diagnosis, and is easy toclinically popularize and use. A circRNA chip is used herein to carry out detecting to obtain disease-specific and abnormally-expressed serum circRNA expression profiles; verifying is performed by means of qRT-PCR; a strict design and evaluation system is provided. It is discovered for the first time that the marker hsa_circ_0001788 in serum is an important biological detection index which causesminimal invasion and has low cost and which is applicable to clinical detection; the marker hsa_circ_0001788 is important to the clinical early diagnosis and differential diagnosis of craniosynostosis; a new idea is provided for the diagnosis of craniosynostosis in molecular level, and it is expected that early diagnostic rate of craniosynostosis is increased.

The invention relates to a method for detecting a circular RNA for bladder cancer screening and an application thereof; in particular, with use of a circular RNA chip, through difference analysis, the circRNA significantly highly expressed in a human bladder cancer tissue is screened and is named as cRNA-ZFR. Compared with a normal tissue, the cRNA-ZFR is significantly highly expressed in the human bladder cancer tissue, and the cRNA-ZFR is further confirmed to have the expression quantity in the human bladder cancer tissue significantly higher than that of the normal tissue in large-sample fluorescent quantitative PCR experiments. The novel cRNA-ZFR provides a new idea for further study of pathogenesis of bladder cancer, and also provides a new target for early screening and prognosis monitoring of bladder cancer.

The invention relates to a general expression framework of artificial circular RNA and application thereof. The general expression framework of artificial circular RNA has multi-microRNA combination inhibition ability. The artificial circular RNA overexpression framework consists of three parts: an upstream sequence, an intermediate sequence and a downstream sequence. The artificial circular RNA overexpression framework designed by the invention can achieve effective overexpression and efficient cyclization of the intermediate sequence in cells to produce the artificial circular RNA with multi-microRNA combination inhibition ability. The invention also relates to application of the general expression framework of the artificial circular RNA and eukaryotic expression plasmids, lentiviruses,adenoviruses, adeno-associated viruses and retroviruses carrying the framework in development and preparation of gene therapy drugs.

The invention discloses a high-flux chip data processing and analysis process control method for circular RNA. The method comprises the steps of generating a self-defined parameter configuration file by a system first and then generating a batch processing executable file corresponding to a data process according to a self-defined parameter file after parameter setting by a user and a high-flux chip data processing process module; and executing the batch processing executable file by the system, implementing data process automation, and finally generating a result report file. Therefore, bio-information analyzers can be efficiently assisted to finish a set of standardized high-flux data analysis process, and even scientific research personnel who do not understand high-flux data analysis can be enabled to finish the high-flux data analysis. In addition, the purposes of optimizing working efficiency of the scientific research personnel and reducing scientific research cost can be achieved. Reliable and diversified circular RNA analysis methods are proposed; and the control method also can be used for high-flux data analysis processes of long-chain non-coding RNA and the like, and is universal in different fields, simple to implement and wide in application range.

The invention belongs to the technical field of biological detection, and particularly relates to a biomarker for detecting mild cognitive impairment (MCI) and/or Alzheimer disease (AD). The biomarkerfor detecting mild cognitive impairment and/or Alzheimer disease is circular RNA, and the circular RNA is ciRS-7. The invention further relates to application of the circular RNA ciRS-7 in detectingmild cognitive impairment and/or Alzheimer disease. By detecting the content of the ciRS-7 in peripheral blood, it is found that the content of the ciRS-7 in the peripheral blood of an MCI or AD patient is higher than that of a normal person, and therefore the ciRS-7 can be used as the biomarker of MCI or AD early diagnosis.

The invention discloses a prediction method for a correlation between circular RNA and a disease based on a gradient enhancement decision-making tree. A circular RNA-disease relationship network is converted into an undirected graph, the circular RNA base sequence similarity, the function annotation semantic similarity and the expression similarity are calculated, and the disease function similarity and the semantic similarity are calculated; a multi-network integration algorithm is adopted for integrating a plurality of kinds of circular RNA similarity networks and conducting weighted averageintegration on a disease similarity network, statistical characteristics of the integrated circular RNA and disease similarity network and the circular RNA-disease relationship network are extracted,and the integrated circular RNA and disease similarity network is converted into an unweighted graph-related characteristic, a circular RNA base sequence characteristic and a circular RNA-disease relationship network implicit vector characteristic; a gradient enhancement decision-making tree learning machine is trained, and a potential circular RNA-disease relationship is predicted. By means of the method, the potential circular RNA-disease relationship can be accurately predicted; and the prediction accuracy of the circular RNA-disease relationship is improved.

The invention provides application of circular DNA (Deoxyribonucleic Acid) in detecting and imaging sense RNA (Ribonucleic Acid) and gene therapy. The circular DNA at least comprises one or more DNA probe fragments complementary to a detection target sense RNA sequence, and is formed by connecting and cyclizing 3' and 5' terminals of ssDNA (single-stranded Deoxyribonucleic Acid). The circular DNAand the detection target sense RNA form heterozygous double strands, and the heterozygous double strands are separated from a carrier, so that (1) the application in detecting and imaging through a corresponding fluorescence signal change is realized; (2) the sense RNA in the heterozygous double strands is enzymically decomposed, so that the aim of gene therapy is achieved.

The invention discloses a method for preparing an exogenous circular RNA and a method for quantitatively determining an endogenous circular RNA. The method for preparing an exogenous circular RNA comprises the following steps: cloning and multiplying an artificially synthesized DAN gene, transcribing the DAN gene into a linear RNA, modifying the tail end of the RNA, and connecting the head and the tail of the RNA to obtain the exogenous circular RNA. Exogenous circular RNA molecules are added to a total RNA according to a specific proportion to serve as a reference gene for real-time quantitative determination of the RNA, and therefore the real-time quantitative determination of the endogenous circular RNA is realized for the first time.

The invention discloses an oligonucleotide composition and a method for detecting hepatitis B virus (HBV) rcDNA (Relaxed Circular deoxyribonucleic acid) and/ or cccDNA (Covalently Closed Circular DNA), a kit and application of the oligonucleotide composition. The oligonucleotide composition comprises first oligonucleotide, second oligonucleotide, or oligonucleotide complementary with the sequenceof the first oligonucleotide and oligonucleotide complementary with the sequence of the second oligonucleotide; the first oligonucleotide is specifically combined with at least one part of continuoussequence of the first sequence, and the second oligonucleotide is specifically combined with at least one part of continuous sequence of the second sequence; in addition, a distance between two continuous sequences is 30-350 nt; the first sequence is a sequence between DR (Direct Repeat) 2-DR1 of the HBV DR sequence of a cross-notch area which contains cccDNA or rcDNA, and the second sequence is asequence after the fifth basic group of the DR1. According to the oligonucleotide composition, the level of HBV rcDNA and/ or cccDNA can be effectively detected without being affected by integrated HBV DNA.

The invention discloses a dye method digital quantitative PCR kit as well as application and an application method thereof. The dye method digital quantitative PCR kit comprises application of circular RNA hsa_circ_0051778 copy number as a detection index in a lung adenocarcinoma and tuberculous pleuritis identification kit according to the circular RNA hsa_circ_0051778 copy number in tuberculouspleuritis being higher than the copy number in the lung adenocarcinoma. The kit comprises a circular RNA hsa_circ_0051778 specific primer, a positive control, a negative control, a special PCR amplification premix solution for dye-method digital PCR as well as sterile deionized water; and the kit is used by virtue of steps of sampling, PCR amplification system preparation, amplification reaction and identification. By adopting the dye method digital quantitative PCR kit, the lung adenocarcinoma and the tuberculous pleuritis can be accurately and rapidly identified. The detection result provesthat the sensitivity of the detection tool can reach about 90 percent or more which is higher than the existing single method used clinically.