38 results about "Palindrome" patented technology

Methods of modifying one or more target nucleic acid sequences using the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR associated (Cas) proteins (CRISPR / Cas) system are disclosed. Methods of introducing one or more exogenous nucleic acid sequences into one or more circular nucleic acid sequences using the CRISPR / Cas system are also disclosed.

Disclosed herein are Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) / CRISPR-associated (Cas) 9-based system related compositions and methods of using said CRISPR / Cas9-based system related compositions for altering gene expression and genome engineering. Also disclosed herein are compositions and methods of using said compositions for altering gene expression and genome engineering in muscle, such as skeletal muscle and cardiac muscle.

A novel immunostimulatory oligonucleotide by which an IFN-inducing activity is enhanced and an inflammatory cytokine-inducing activity is reduced, and a pharmaceutical containing the same, and an application thereof are provided. That is, the present invention provides the immunostimulatory oligonucleotide composed of a base sequence represented by a formula: 5′-(G)MPXCGYQ(G)N-3′ (X and Y are mutually independent and represent an arbitrary sequence which has a length of 0 to 10 nucleotides and does not contain 4 or more consecutive G residues, and a length of X+Y is 6 to 20 nucleotides; XCGY contains a palindrome sequence having a length of at least 8 nucleotides and has a length of 8 to 22 nucleotides; P and Q are mutually independent and represent one nucleotide other than G; M represents an integer of 6 to 10 and N represents an integer of 0 to 3) wherein a full length thereof is 16 to 37 nucleotides (except for an oligonucleotide composed of a basesequence represented by SEQ ID NO:5), the pharmaceutical application thereof.

A novel immunostimulatory oligonucleotide by which an IFN-inducing activity is enhanced and an inflammatory cytokine-inducing activity is reduced, and a pharmaceutical containing the same, and an application thereof are provided. That is, the present invention provides the immunostimulatory oligonucleotide composed of a base sequence represented by a formula: 5′-(G)MPXCGYQ(G)N-3′ (SEQ ID NO: 118) (X and Y are mutually independent and represent an arbitrary sequence which has a length of 0 to 10 nucleotides and does not contain 4 or more consecutive G residues, and a length of X+Y is 6 to 20 nucleotides; XCGY contains a palindrome sequence having a length of at least 8 nucleotides and has a length of 8 to 22 nucleotides; P and Q are mutually independent and represent one nucleotide other than G; M represents an integer of 6 to 10 and N represents an integer of 0 to 3) wherein a full length thereof is 16 to 37 nucleotides (except for an oligonucleotide composed of a base sequence represented by SEQ ID NO:5), the pharmaceutical application thereof.

The invention provides a system and method for synthesizing polynucleotides by solid phase assembly oligonucleotide precursors, in accordance with the method, a polynucleotide is partitioned into an ordered set of subunits, wherein each subunit is assembled in a single reaction from a subset of oligonucleotide precursors that uniquely anneal together to produce the subunit. The subunits are then assembled to form the desired polynucleotide. An important feature of the invention is the selection of subunits that are free of undesired sequence elements, such as palindromes, repetitive sequences, and the like, which would result in more than one subunit product alter ligating a pool of oligonucleotide precursors.

The invention discloses a method for improving the yield of vernine from bacillus amyloliquefaciens. The method comprises the following steps: by taking thermo-sensitive type plasmid pKS2 as a carrier, removing a promoter partial segment of a purine operon (purEKBCSQLFMNHD) through adopting a homologous recombination method, wherein the preferable sequence segment is -193bp to -29bp segment of the purE-deleted gene, and the repression effect of purR (repressor protein of the purine operon) and the transcription attenuation action caused by the palindrome structure of the segment of a strain with the segment deleted are removed; and desensitizing transamidase gene of purF coded phosphoribosyl pyrophosphate (PRPP) through point mutation, wherein preferably, leucine at 217th site is mutated into isoleucine (L217I), and the feedback inhibition from products adenosine and vernine can be removed. The production capability of the transformed genetically engineered bacterium vernine is obviously improved.

The invention relates to a fluorescent probe for fluorescent quantitative PCR reactions. The fluorescent probe is characterized in that the probe comprises an oligonucleotide sequence, the 5' terminus is a specific oligonucleotide sequence with an amplified target sequence and modifies the fluorescence groups; the 3' terminus is a nucleotide sequence, forms a reversely-complimentary palindrome structure with the 5' terminus, and modifies the fluorescence quenching groups. Under certain conditions, the probe can form an intramolecular secondary composition, and the fluorescent groups get close to the quenching groups so as to quench the fluorescence signals released by the fluorescence groups. Compared to the common fluorescent probe, the provided fluorescent probe has the advantages of high specificity, high sensitivity, and low fluorescent quenching efficiency, thus the signal to noise ratio (SNR) of the reactions is higher, and the provided fluorescent probe can be applied to realtime fluorescence quantitative PCR detection reactions and all end point fluorescence detection reactions.

The present invention relates to enzymes, compositions and methods for catalyzing asymmetric recombination of non-palindromic recombination sites in a cell free system, in isolated cells or in living organisms. The enzymes and methods of the invention are suitable for mediating specific recombinations between DNA sequences comprising specific recombination sites without being limited to strict palindromic symmetry within each recombination site.

The invention belongs to the field of nucleic acid editing and particularly relates to the technical field of clustered regularly interspaced short palindromic repeat (CRISPR). Specifically, the invention provides a gRNA and the gRNA can be used for improving efficiency of a Cas enzyme.

The present invention generally relates to libraries, compositions, methods, applications, kits and screens used in functional genomics that focus on gene function in a cell and that may use vector systems and other aspects related to Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas systems and components thereof. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for utilizing the CRISPR-Cas system.

The present invention provides a method for rapidly detecting the genome-wide presence of palindrome formation. The method has demonstrated that somatic palindromes occur frequently and are widespread in human cancers. Individual tumor types have a characteristic non-random distribution of palindromes in their genome and a small subset of the palindromic loci are associate with gene amplification. The disclosed method can be used to define the plurality of genomic DNA palindromes associated with various tumor types and can provide methods for the classification of tumors, and the diagnosis, early detection of cancer as well as the monitoring of disease recurrence and assessment of residual disease.

The invention provides gRNA targeted to Rett syndrome-related mutation gene RNA and provides a detection method and a detection kit for human Rett syndrome-related mutation on the basis of a clusteredregularly interspaced short palindromic repeats (CRISPR)-C2c2 system. In the detection method, by means of advantages of the gRNA targetedly-recognizing a transcription product RNA (target RNA sequence) of the Rett syndrome-related mutation gene, when the CRISPR-C2c2 composite detects the target RNA sequence, the composite can cut the report RNA having a detection label and release a detectable signal, so that when the CRISPR-C2c2 system is applied to detection for the Rett syndrome-related mutation gene, high sensitivity and accuracy are achieved. The detection method and the detection kit have huge commercial application potential value.

A perfect palindrome operator sequence-based protein expression system is provided. The expression system comprises a promoter; and a perfect palindrome operator sequence, wherein the promoter is not T7. The expression system is preferably employed for the production of recombinant proteins by fermentation.

The invention provides a method for detecting a tumor biomarker by using a palindrome lock-type probe. A palindrome-sequence non-fluorescent mark lock-type RCA probe is designed. The probe is composedof a target identification part and a locus cutting part. The target identification part and target miRNA can completely complement each other and is located at a 3' end and a 5' end of the lock-typeprobe; a semi-identification locus of restriction enzyme Nt.AlWI is fused with a palindrome alkali segment. The probe is combined with nicking enzyme, an N-RCA concept is provided, a double-directionchain replacement reaction (D-SDA) is conducted for combination, and the probe is used for detecting the tumor marker miRNA. By means of the method, let-7a miRNA can be efficiently detected through amplification, and the probe is economical and practical and is expected to be applied to development of clinical diagnostic kits.

The invention relates to a Leon-RC compression method for genome sequencing data. The method mainly aims to improve the steps of constructing an anchor point dictionary by an LEON algorithm. The method includes the following steps that: (1) a short-read is divided into a plurality of Kmers; (2) one Kmer is selected, the Kmer values of the direct repetition, mirror repetition, inversion repetition,and complementary palindrome of the Kmer are calculated, the four values are compared with one another, so that a minimum Kmer value can be obtained; (3) the minimum Kmer value is inputted into a Bloom filter for matching search, a Solid Kmer is stored in the Bloom filter, and whether the minimum Kmer value exists in the Solid Kmer is judged; if the minimum Kmer value exists in the Solid Kmer, the minimum Kmer value is added to the anchor point dictionary, and search is ended; if the minimum Kmer value does not exist in the Solid Kmer, the next Kmer is obtained, and the step (2) and step (3)are repeated; (4) if the minimum Kmer values of all the Kmer do not exist in the Solid Kmer, it is indicated that no anchor point exists in the short-read; and (5) the anchor point dictionary is constructed through the step (1) to (4).

The invention discloses a pseudo attP site in the swine genome and applications thereof, belonging to the technical field of biotechnology and biomedical engineering. According to the invention, a pseudo attP site is isolated from the swine genome, and the pseudo attP site has a sequence similarity of 40% with the wild type attP site and has the characteristic of palindromic sequence. The pseudo attP site can be recognized by Streptomycete bacteriophage phi C 31 intergrase, and thereby exogenous genes having attP site can be mediated to integrate in the site in relatively high efficiency. According to protein content tests, the expression level of EGFP gene maintained by the pseudo attP site is 50 times that by the genome without exogenous genes. The invention provides a new technical scheme for targeted integration and high efficiency expression for swine transgene, and a new strategy for the research of swine functional genome, so the invention has important science value and broad application prospect.

Provided herein are compositions, methods, kits, and viral particles for treating a disease or disorder associated with a deep intronic mutation using an engineered, non-naturally occurring Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associated (Cas) (CRISPR-Cas) system. In some aspects, provided herein is a self-limiting CRISPER-Cas system.

The invention provides a PCR primer and an application thereof in DNA fragment connection. The PCR primer comprises a palindrome sequence segment and any PCR primer sequence segment, the 3'end of thepalindrome sequence segment is connected with the 5'end of the any PCR primer sequence segment, the nucleotide of the 3' end of the palindrome sequence segment is U, and the any PCR primer sequence segment is complementarily paired with the 3'end of a pre-amplified DNA template. The primer provided by the embodiment of the invention can be used for constructing a sequencing library; in the construction process, the primer provided by the embodiment of the invention is used as a PCR amplification primer based on DNA, the two ends of an obtained amplification product DNA double strand have a palindrome sequence structure, and the 3'end of the palindrome sequence structure has nucleotide U, so that a foundation is laid for subsequent interconnection of different amplification product double strand DNAs.

The invention relates to the technical field of biosensors, in particular to a palindrome molecular beacon-based biosensor for detecting adenosine triphosphate (ATP). The palindrome molecular beacon-based biosensor comprises a palindrome molecular beacon MB, an ATP adaptor (split adaptor probes AP1 and AP2), target object ATP, Bst DNA polymeras, UDG, endonuclease IV and a buffer liquid. Accordingto specificity identification of an aptamer and the target object, fragment tail sequences of the two split adaptors get close to each other, a hairpin structure of the palindrome molecular beacon MBis opened to generate fluorescence, at the moment, a palindrome sequence locked in advance is released, intermolecular hybridization is performed, so that the spontaneous polymerization, repair, internal cut and circulation process is triggered, the amplification of a fluorescent signal is achieved, and an aptamer biosensor is built. The sensor reaction only needs a step, thus, the biosensor has the advantages of rapid detection speed, low cost, low detection limit, high specificity and the like and is simple and convenient to operate.

The invention provides a system and method for synthesizing polynucleotides by solid phase assembly oligonucleotide precursors, in accordance with the method, a polynucleotide is partitioned into an ordered set of subunits, wherein each subunit is assembled in a single reaction from a subset of oligonucleotide precursors that uniquely anneal together to produce the subunit. The subunits are then assembled to form the desired polynucleotide. An important feature of the invention is the selection of subunits that are free of undesired sequence elements, such as palindromes, repetitive sequences, and the like, which would result in more than one subunit product alter ligating a pool of oligonucleotide precursors.

Belonging to the technical field of genetic engineering, the invention discloses a repetitive extragenic palindromic sequence for improving exogenous gene mRNA and application thereof. The repetitiveextragenic palindromic sequence and a spacer region sequence thereof are shown as SEQ ID NO, 1, the signal peptide can improve the stability of mRNA by changing the secondary structure, so that the extracellular enzyme activity of the target protein cyclodextrin glycosyltransferase can be improved by 14%, the recombinant escherichia coli modified by the signal peptide has strengthened extracellular protein production capacity, therefore the repetitive extragenic palindromic sequence can be used for escherichia coli to express exogenous protein.

The invention provides a PCR primer and an application thereof in DNA fragment connection. The PCR primer comprises a palindrome sequence segment and any PCR primer sequence segment, the 3'end of thepalindrome sequence segment is connected with the 5'end of the any PCR primer sequence segment, the nucleotide of the 3' end of the palindrome sequence segment is U, and the any PCR primer sequence segment is complementarily paired with the 3'end of a pre-amplified DNA template. The primer provided by the embodiment of the invention can be used for constructing a sequencing library; in the construction process, the primer provided by the embodiment of the invention is used as a PCR amplification primer based on DNA, the two ends of an obtained amplification product DNA double strand have a palindrome sequence structure, and the 3'end of the palindrome sequence structure has nucleotide U, so that a foundation is laid for subsequent interconnection of different amplification product double strand DNAs.

The invention discloses a mitochondrion tRNA (transfer ribonucleic acid)<Thr> 15909A>G mutation detection method and a kit for detecting mitochondrion tRNA<Thr> 15909A>G mutation. The method includes DNA (deoxyribonucleic acid) extraction, specific primer design, specific band PCR (polymerase chain reaction) amplification, specific restriction endonuclease HpyCH4V enzyme digestion, agarose gel electrophoresis identification and the like. After being subjected to specific PCR amplification, a DNA sample positioned at a tRNA<Thr> 15909A>G mutation position possibly comprises a TGCA palindrome structure which can be specifically recognized by specific restriction endonuclease HpyCH4V, then two small fragments are obtained by rapid enzyme digestion, and two bands are obtained after agarose gel electrophoresis. The mitochondrion tRNA<Thr> 15909A>G mutation detection method and the kit have the advantages that requirements on required detection instruments and required detection reagent cost are low, and according to the required method, simplicity and stability in operation, easiness in rapid mastering, accuracy, good specificity and the like are achieved; the method and the kit are beneficial to establishment of perfect screening technology for tRNA<Thr> mutation and can be widely popularized to clinical detection.

The invention relates to a microfluidic-technology-based CTC protein typing kit. The kit comprises: (1) capture antibody compounds aiming at different CTCs marker protein, each capture antibody compound being made from a capture antibody in specific binding to corresponding marker protein and an aptamer, wherein the capture antibody and aptamer are connected through a cuttable material which is deoxyribonucleotide double strands having a palindrome structure and two or more than two restriction enzyme sites; and (2) a microfluidic chip, the surface of which is coupled to a cell capture assembly. The cell capture assembly made from a microcolumn is fixed at the surface of the microfluidic chip, and a specific conjugate capable of bonding to the aptamer modifies one end of the microcolumn. Through the microfluidic technology, efficient and fast automatic detection and typing of CTCs is achieved, the kit is simple to operate, false-negative and false-positive results due to man-made operational factors are reduced, and the sensitivity and accuracy of detection are improved.

The invention discloses an in-situ hybridization detection kit comprising a hybridization probe and a marker, and also discloses a method using the kit to carry out in-situ hybridization detection on mRNA (messenger ribonucleic acid) of a palindrome gene (FUBP1) closely related to early stage pathology evolution of brain glioma. The method comprises the following steps of: (1) under the condition that the hybridization probe and a target sequence can form a stable hybridization compound, contacting the RNA to be detected in a substrate with the hybridization probe to form the hybridization complex; and (2) detecting the hybridization complex. The kit and the detection method disclosed by the invention can detect the expression quantity of gene in mRNA level, and can detect pathological changes earlier than image medicine and current biochemical detection indices, thus being capable of realizing real mRNA-level screening at the early stage of canceration; and meanwhile, the detection method disclosed by the invention is simple and convenient, low in cost and convenient to be popularized and applied in county and district hospitals.

Disclosed herein are Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) / CRISPR-associated (Cas) 9-based epigenome modifier compositions for epigenomic modification of a SNCA gene and methods of use thereof.

The invention relates to a Leon-RC compression method for genome sequencing data, which mainly improves the steps of constructing an anchor dictionary by the LEON algorithm, including the following steps: (1) dividing short reads into multiple Kmers; (2) selecting one Kmer, calculate the Kmer value of its direct repetition, mirror repetition, inverted repetition, and complementary palindrome, compare these four values, and obtain the smallest Kmer value; (3) Put the smallest Kmer value into the Bloom filter for matching Search, Solid Kmer is stored in the Bloom filter, and judge whether there is a minimum Kmer value in Solid Kmer; if it exists, add the minimum Kmer value to the anchor dictionary and end the search; if not, obtain the next One Kmer, repeat steps (2), (3); (4) If the minimum Kmer value of all Kmers does not exist in Solid Kmer, it means that there is no anchor point for this short read; (5) Pass step (1) ~(4) Construct the anchor dictionary.