17 results about "Dna breaks" patented technology

Compositions and methods are provided for high-efficiency large scale manipulation of genomic regions and chromosomal engineering of plant genomes. Enhancement of chromosomal modification includes the provision of one or more linker oligonucleotides complementary to the distinct chromosomal ends following double strand breaks, increasing the temperature effect and / or recurrent cutting by an endonuclease to facilitate higher chromosomal modification including large segment translocations and recombination. Site-specific directed DNA breaks under one or more of the experimental conditions (including CRISPR-Cas systems) disclosed enhance targeted recombination frequencies, crossover efficiency and movement of large chromosomal segments in crop plant cells.

Cancer poses a significant threat to human health, and currently there is no particularly effective treatment. The primary reason is the lack of a highly effective method to specifically kill cancer cells while minimizing impact on normal cells. DNA breaks, if left unrepaired, lead to cell death—this is the fundamental principle of radiotherapy—but radiotherapy also causes DNA damage in normal cells. This invention proposes a method for specifically killing cancer cells. It utilizes gene editing technology to create DNA breaks unique to cancer cells (these DNA cut points are absent in normal cells), simultaneously combining this with DNA damage repair inhibitors to suppress DNA repair in cancer cells, leading to specific cancer cell death. This invention provides a new approach to precision cancer treatment.

The invention relates to a device and a method for purifying a DNA-containing sample, the device comprises: a sample chamber used for containing a DNA-containing sample to be treated, a first semipermeable membrane arranged at a first end and a second semipermeable membrane arranged at a second end, the first semipermeable membrane and the second semipermeable membrane being in membrane structures, and the first semipermeable membrane and the second semipermeable membrane being in a direction from the inner side to the outer side of the sample chamber, the first end and the second end can pass through a non-target isolate but cannot pass through a target isolate, an electrophoretic field is formed between the first end and the second end, the target isolate is DNA, and the non-target isolate comprises charged impurities; the first ends of the first liquid storage device and the second liquid storage device are sealed in the first liquid storage device, the second ends of the first liquid storage device and the second liquid storage device are sealed in the second liquid storage device, and the first liquid storage device and the second liquid storage device are used for containing an electrophoretic buffer solution; an electrode on one side of the electrode assembly extends into the first liquid storage device, an electrode on the other side of the electrode assembly extends into the second liquid storage device, a loop is formed, and a sample containing DNA is subjected to electrophoresis. By means of electrophoresis, impurities and DNA are migrated together, the DNA is intercepted through the semi-permeable membrane, the impurities can be removed through the semi-permeable membrane, and the purposes of removing the impurities such as protein and polysaccharide and purifying the DNA are achieved. Due to the fact that a mild impurity removal mode is adopted, operation such as violent vibration is avoided, the possibility that DNA breaks in the purification process is reduced, and the purposes of purifying DNA and keeping the length of the DNA at the same time are achieved.

The application discloses a dCas9-based c-NHEJ site-directed inhibition system and application in improving HR-mediated gene editing efficiency, and the system comprises the following elements: (1) element 1: a nuclease for site-directed induction of DSB in eukaryotic cells and accompanying elements thereof; (2) element 2: dCas9-sgRNA for binding to the end of DNA break; (3) element 3: a homologous template for HR-mediated gene editing. The system synergistically improves gene editing efficiency. The application provides site-directed and efficient inhibition of the c-NHEJ repair pathway, site-directed and efficient improvement of the HR repair efficiency, effective improvement of the HR-mediated CRISPR gene editing efficiency, and reduction of off-target effects.

This specification discloses the use of alternating electric fields and DNA-dependent PK inhibitors for cancer treatment. This specification discloses the use of alternating electric fields and DNA-dependent PK inhibitors for inducing cell death. This specification discloses the use of alternating electric fields and DNA-dependent PK inhibitors to inhibit DNA repair in cancer cells having DNA strand breaks (e.g., double-strand DNA breaks). This specification discloses the use of alternating electric fields and DNA-dependent PK inhibitors to enhance the effectiveness of radiotherapy.

Compositions and methods are provided for genetically modifying a cell to introduce both a gene knockin and a gene knockout. The subject methods use a donor polynucleotide comprising a gene knockin sequence and a knockout guide RNA sequence. Transcription of the donor polynucleotide, after integration of the donor polynucleotide into the genome, produces a mature mRNA sequence comprising the knockout guide RNA. An RNA-guided nuclease is used to excise the knockout guide RNA from the mRNA transcript, which then guides the RNA-guided nuclease to a second target locus where the RNA-guided nuclease creates a double stranded DNA break, resulting in gene knockout at the second target locus. Only cells with the gene knockin are able to express the mRNA containing the knockout guide RNA so that the gene knockin at the first genomic target locus and the subsequent gene knockout at the second genomic target locus are linked.

To provide a system and process for identifying and characterizing double-stranded DNA break repair sites that are based on biological information, and have improved accuracy.SOLUTION: A process is configured to: receive sample sequence data including a plurality of sequences; analyze and merge the sample sequence data; when a single-stranded or double-stranded DNA oligonucleotide donor is provided, output a target site sequence including a prediction result of a repair event; bine the merged sequence with the target site sequence or an arbitrary target prediction result; output a target-read realignment; realign a binned target-read realignment with a target site; generate a final alignment; analyze the final alignment; identify and quantify mutations within a pre-defined sequence distance window deriving from a canonical enzyme specific cut site; and output the final alignment, and data on results of the analysis and quantification as tables or graphics.SELECTED DRAWING: None

The present application belongs to the technical field of biological medicine, and particularly relates to a kind of targeting REXO4 liquid-liquid phase separation interference polypeptide and its application.The interference polypeptide of the present application includes targeting peptide, the targeting peptide is specifically combined with REXO4 protein liquid-liquid phase separation driving domain, and the liquid-liquid phase separation of REXO4 protein is interfered;Wherein, the REXO4 protein liquid-liquid phase separation driving domain is REXO4 protein N-terminal disordered domain, and the amino acid sequence of the REXO4 protein N-terminal disordered domain is shown as sequence 1.The interference polypeptide of the present application can be specifically combined with REXO4 protein liquid-liquid phase separation driving domain, interfere with and destroy the functional LLPS of REXO4, cause the DNA damage repair efficiency in tumor cell to be significantly reduced, lead to the DNA break caused by ionizing radiation cannot be repaired in time, finally enhance the sensitivity of tumor cell to ionizing radiation, realize the effect of radiotherapy sensitization.

The utility model provides a bottom support for a DNA (Deoxyribose Nucleic Acid) breaking instrument, which belongs to the technical field of DNA breaking instruments, comprises a motor kinetic energy device for driving a test tube for containing a DNA sample to rotate, and is characterized by further comprising a platform plate for supporting and a DNA mounting module for normatively placing the test tube, a central through hole is formed in the middle of the platform plate, and the DNA installation module is placed and installed in the central through hole; the platform plate is further provided with a notch used for containing the motor kinetic energy device. A driving lever for driving the DNA mounting module to rotate is fixedly mounted on an output shaft of the motor kinetic energy device, and contacts are arranged at the two ends of the bottom of the driving lever; the bottom platform of the DNA breaking instrument can solve the problems that when an existing DNA breaking instrument is used, a tray for containing DNA sample test tubes shakes, and the whole tray is difficult to take and place.

Described herein is a system and process for identifying and characterizing double-stranded DNA break repair sites that are based on biological information and have improved accuracy. Also described is a sequence alignment process that uses biological data to inform the alignment matrix for position specific alignment scoring, resulting in the identification of noncanonical target sites.

B cells with an altered function. FIG. 1A Rearrangement at the Ig locus n - 250-1,000) VH1 VHP DH1-12 T FEu DNA-PKcs RAG1 / 2 VIDN Excised Ku70, Ku80 NHEJ recombination intervening DNA ARTEMIS XROC4 DNA Ligase N V DJ Cp C6 OB Cr1 0120 area Ce Cex 3' RR Eu Su Se At S regions: DNA breaks AD Class-switch DNA repair recombination IgM V D JCu Transcription Recombined CH locus V DJ Ce Ca 3' RR Cy2a Cu Eµ C8 Excised Cy2b intervening DNA IgE V DJCe Cy3 Cyl 20 26 20 47 61 19 J un 2 02 6 A B S T R A C T 1 9 J u n 2 0 2 6 2 0 2 6 2 0 4 7 6 1 F I G . 1 A R e a r r a n g e m e n t a t t h e I g l o c u s V H 1 V H n D H 1 - 1 2 7 F E u r D N A - P K c s R A G 1 / 2 V I D N N H E J V D J C p C 6 O B 0 1 2 0 O R D C e E u S u S e A D r e c o m b i n a t i o n I g M V D J C u R e c o m b i n e d V D J C e C a 3 ' R R C y 2 a C u E u r C 8 E x c i s e d C y 2 b I g E V D J C e C y 3 C y l

To provide combination therapy comprising administering a radioimmunoconjugate and an inhibitor of DNA damage response.SOLUTION: The present disclosure encompasses the insight that combining inhibition of DNA damage-repair mechanisms with a therapy that targets DNA breaks to cancer cells may provide a less toxic therapy with improved efficacy. Radioactive decay can cause direct physical damage (such as single or double-stranded DNA breaks) or indirect damage (such as by-stander or crossfire effects) to the biomolecules that constitute a cell. The disclosure combines a radioimmunoconjugate targeting cancer cells with an inhibition of DNA damage-repair to treat or ameliorate cancer.SELECTED DRAWING: None

The invention discloses a deep learning-based CRISPR / Cas9 mediated DNA double-strand fracture repair result prediction method, which comprises the following steps: (1) constructing a data set, and obtaining a DNA sequence data set from a public database; (2) constructing a Token dictionary which is used for coding the DNA sequence into a digital sequence; and (3) constructing an input sequence, scanning the DNA sequence to obtain 1-mer and 3-mer subsequences, splicing the 1-mer and 3-mer subsequences, and finally encoding the 1-mer and 3-mer subsequences into a digital sequence for model input through a Token dictionary. And (4) designing an ECnnIndel method based on deep learning, and predicting a CRISPR / Cas9 editing result. And (5) encoding a DNA sequence to be detected into a digital sequence through the steps, inputting the digital sequence into the trained ECnnIndel model, and analyzing the generalization performance of the ECnnIndel. By adopting the method, the defects of insufficient equipment resources, low precision and the like are effectively overcome, and the ECnnIndel promotes the research progress of a computer-aided double-stranded DNA fracture repair mechanism.

The invention relates to the technical field of genetic engineering and bioinformatics, in particular to a method for creating a new gene in a living body on the premise of no artificial DNA template and application. The method is characterized in that DNA fractures are simultaneously generated at at least two different specific positions in the genome of an organism, the specific positions are genomic sites capable of segmenting different gene elements or different protein structural domains, and the DNA fractures are connected with one another by means of non-homologous end connection (NHEJ) or homologous repair. And generating a new combination of the different gene elements or different protein structural domains, which is different from the original genome sequence, so as to form a new gene. The new gene provided by the invention can change the growth, development, resistance, yield and other characters of organisms, and has important application value.

In some aspects, this disclosure provides a high throughput method of identifying highly active and / or highly specific guide RNAs.

The utility model discloses an ultrasonic DNA breaking instrument which comprises a shell, a telescopic arm is fixedly connected in the shell, an amplitude-change pole is arranged at the top end of the telescopic arm, a control screen is arranged at the top of the shell, a sliding rail is further arranged in the shell in the vertical direction, the sliding rail is connected with a moving rod, the sliding rail and the moving rod are symmetrically arranged, and a rotating shaft is arranged at the bottom of the shell. The ultrasonic DNA breaking instrument is low in manufacturing cost, efficient, accurate, easy and convenient to operate, high in compatibility and capable of being suitable for different containers such as test tubes and beakers, the amplitude-change pole can be adjusted according to the sample capacity, and the tedious operation of manual replacement is avoided.

Combination therapy comprising administering a PSMA-targeted radiopharmaceutical and one or more checkpoint inhibitors. The present disclosure encompasses the insight that combining the inhibition of checkpoint proteins with a treatment that targets damage to cancer cells may provide a less toxic treatment with improved efficacy. Radioactive decay can cause direct physical damage (such as single- or double-stranded DNA breaks) or indirect damage (such as bystander or crossfire effects) to the biomolecules that make up cells. Drugs that deliver radionuclides to cancer cells, i.e., radiopharmaceuticals, provide a mechanism for generating DNA damage with anticancer therapeutic effects.