41 results about "Dna breakage" patented technology

The invention provides for a method of producing a mutant somatic human cell line of cells comprising a genomic mutation of interest (MOI) at a predefined genomic site of interest (GOI) in close proximity to a genomic target site, which comprises: a) providing a guide RNA (gRNA) comprising a tracrRNA in conjunction with crRNA including an oligonucleotide sequence that hybridizes with the target site; b) providing an RNA-guided endonuclease which catalyzes the DNA break at the target site upon hybridizing with the gRNA; c) introducing the gRNA into the cells in the presence of the endonuclease to obtain a repertoire of cells comprising a variety of genomic mutations at the target site; d) selecting a cell from said repertoire which comprises a MOI; wherein the cell is haploid for the genomic locus of the target site; and e) expanding the cell to obtain the mutant cell line. The invention further provides for a mutant human somatic cell line obtainable by such method; and libraries of mutant human somatic cell lines of isogenic cells with a variety of genomic mutations at different predefined genomic target sites.

The invention discloses a high-temperature combined method for extracting genomic DNA of fish enteric microorganisms. The high-temperature combined method includes the steps that firstly, the walls of various fish enteric microorganisms are properly broken through a lysozyme method and an ultrasonic method at the same time, wherein the ultrasonic physical wall breaking condition ranges from 150 w to 250 w, ultrasonic treatment is carried out for 2-3 s and then carried out again after an interval of 5 s, and the process is repeated 50-60 times; secondly, the sample obtained after ultrasonic treatment and lysozyme are jointly incubated for 30 min in a water-bath constant-temperature oscillator at the temperature of 55-65 DEG C, and the mixture and RNase A are jointly incubated at the temperature of 27-32 DEG C. According to the method, the walls of the various microorganisms are properly broken under the proper and mild ultrasonic condition, and the genomic DNA is prevented from being broken and degraded. As the specific temperature range for incubating the lysozyme and the sample and the specific temperature range for incubating the RNase A and the sample are set, the splitting decomposition effect of the lysozyme can be improved, and it can be avoided that the genomic DNA is degraded by DNA enzymes in tissues and cells. Operation is easy, the extracted genomic DNA is complete and low in degradation rate, and the fish enteric microorganism structure can be truly reflected by the types of the microorganisms identified according to the extracted DNA.

Methods of editing a target genomic region(s), methods of repairing of a DNA break via a HDR pathway, methods of inhibiting or suppressing repair of a DNA break via a NHEJ pathway, and methods of modifying expression of a gene(s) or protein(s) comprise administering to one or more cells that include one or more target genomic regions, a genome editing system and a DNA protein-kinase (DNAPK) inhibitor disclosed herein. Kits and compositions for editing a target gene comprise a genome editing system and a DNAPK inhibitor disclosed herein.

The invention relates to compositions and methods for interfering with the DNA repair of double strand breaks (DSBs). The invention discloses novel double-stranded nucleic acid molecules that act as baits and hijack the holocomplex of enzymes responsible of DNA DSB sensing, signaling and / or repair pathways, in particular the non homologous end joining (NHEJ) pathway of DSB repair. The invention discloses the use of these molecules as adjuvant compositions to be used in association with a DNA breaking treatment, particularly radiotherapy or chemotherapy, in combination with a pharmaceutically acceptable carrier, in an efficient amount to be introduced in the tumor cell nuclei in order to neutralize transiently their DNA repair capacity and trigger their death.

The invention provides sgRNA, a CREBRF point mutation-type Bama miniature pig constructed by the sgRNA, and application. The sgRNA comprises a nucleic acid sequence as shown in SEQ ID NO: 1 or SEQ IDNO: 2. According to the invention, a scheme of combining CRISPR / Cas9 with homologous recombination is adopted, point mutation of CREBRF gene is carried out, the designed gRNA has specific binding property on a target site near the 1370th site of the CEREBRF gene, the CRISPR / Cas9 can be guided to carry out specific recognition cutting near the target site, double-stranded DNA breakage is formed, homologous recombination repair with exogenous single-stranded DNA as a template is promoted, and therefore the point mutation is introduced. The mutant Bama miniature pig prepared by the method has a typical obesity phenotype and can be used as a model organism to be applied to the field of research and development of medicines for treating obesity and diabetes.

The invention relates to the field of fish sperm preservation solutions, in particular to a pseudobagrus sperm preservation solution for solving the problem of pseudobagrus large scale reproduction and a preparation method of the pseudobagrus sperm preservation solution. The sperm preservation solution is prepared from sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium bicarbonate, glucose and penicillin and is prepared by weighing the raw materials, adding the raw materials in a plastic bottle or a glass bottle, and preparing into a solution with distilled water. The prepared sperm preservation solution is subjected to sperm chromatin detection with a flow cytometer, spermatids are circled on an FSC / SSC graph, and a door is set on an FL1(FITC) / FL3(ECD) scatter diagram for analysis; and a DNA fragment index at is calculated through a formula to evaluate sperm DNA quality. The sperm preservation solution provided by the invention can be preserved for 10 days in a refrigeration tank at the temperature of 4 DEG C, 98 percent or more of sperms keep exuberant vigor and can still be taken as sperms for semination, and the fertilization rate is 95 percent orabove.

The present invention provides a simple and rapid method for preparing purified transposase complexes that are highly suited for fragmenting DNA. The method includes forming transposase complexes with oligonucleotide adapters in cell lysate, then purifying the complexes from the other substance in the cell lysate. Purification is accomplished using a specific binding pair, in which one member of the pair is bound to an oligonucleotide adapter of the complex and the other member of the pair is bound to a solid substrate. The bound complexes can be immediately used in DNA fragmentation reactions to produce solid substrate-bound DNA fragments, which can be used for any number of purposes, including as templates for amplification and sequencing.

The invention discloses a novel tetracyclonaphthooxazole derivative. The structure of the novel tetracyclonaphthooxazole derivative is represented by general formula (I) shown in the specification. The invention also discloses a preparation method of the tetracyclonaphthooxazole derivative. The method allows the tetracyclonaphthooxazole derivative to be innovatively synthesized through using a carbon hydrogen activation serial reaction with amino substituted quinone as a reaction substrate. The method has the advantages of simplicity, easy implementation and high efficiency. The synthetic tetracyclonaphthooxazole derivative has a similar structure with a traditional DNA clastogen, can inhibit cell proliferation and growth, and has antibacterial, antifungal, anti-HIV and antitumor bioactivity. The invention also discloses an application of above compounds as a cell proliferation and growth inhibitor, and an application of the compounds in the preparation of antitumor medicines.

The invention relates to a method and a system for sorting sperm samples according to different levels of DNA fragmentation and methods of using populations with low levels of DNA fragmentation to improve fertility and success rates of assisted reproductive procedures, including artificial insemination, in vitro fertilization, intracytoplasmic injection, and other related techniques.

The present invention relates generally to a method of amplifying a nucleic acid region of interest and, more particularly, to a method of amplifying a nucleic acid region of interest using a PCR method designed to minimise the generation of amplicons from primers which have bound to nucleic acid regions other than the specific region of interest. The method of the present invention is based on the determination that by rendering inefficient the functionality of either the forward primer or the reverse primer, the rate of amplification of irrelevant nucleic acid regions can be reduced relative to amplification of the region of interest. The provision of a selective means of amplifying a nucleic acid region of interest is useful in a range of applications including, but not limited to, the diagnosis and / or monitoring of disease conditions which are characterised by specific gene sequences, the characterisation or analysis of gene regions of interest, the identification or characterisation of DNA breakpoint regions and the isolation of gene sequences of interest where only the nucleotide sequence at one end of the gene sequence of interest is known.

Provided is a genome shuffling technology suited to creating a useful next-generation plant body.Genome shuffling is performed using a polypeptide comprising a first polypeptide region for double-stranded DNA breakage activity and a second polypeptide region for intertissue migration activity.

The invention relates to a method for detecting a PAS (Poly A Site) of an RNA (Ribose Nucleic Acid), and provides a method for analyzing APA (Alternative Polyadenylation) based on 3T-seq (TT-mRNA Terminal Sequencing) within a whole-genome range. The method for analyzing APA comprises the following steps: preparing magnetic beads containing oligo dT, i.e., mixing an oligo dT primer modified by 5' end biotin and the magnetic beads with streptavidin; mixing the magnetic beads with total RNA and screening RNA containing Poly A; performing reverse transcription, synthesizing the second strand of a double-stranded cDNA and breaking the double-stranded cDNA; removing the Poly A structure; after terminal modification, connecting a sequencing linker; and recovering libraries. According to the method for analyzing APA, RNA horizontal operation is reduced, a Poly A sequence is horizontally removed from the DNA, and the influence of the Poly structure on sequencing is eliminated; double-stranded DNA breaking enzyme is selected for treatment, and on the premise that the quality of the libraries is not affected, the DNA is broken horizontally; the experiment process is simplified, the experiment difficulty and cost are lowered and the method for analyzing APA is wide in application range.

The invention discloses a high-temperature combined method for extracting genomic DNA of fish enteric microorganisms. The high-temperature combined method includes the steps that firstly, the walls of various fish enteric microorganisms are properly broken through a lysozyme method and an ultrasonic method at the same time, wherein the ultrasonic physical wall breaking condition ranges from 150 w to 250 w, ultrasonic treatment is carried out for 2-3 s and then carried out again after an interval of 5 s, and the process is repeated 50-60 times; secondly, the sample obtained after ultrasonic treatment and lysozyme are jointly incubated for 30 min in a water-bath constant-temperature oscillator at the temperature of 55-65 DEG C, and the mixture and RNase A are jointly incubated at the temperature of 27-32 DEG C. According to the method, the walls of the various microorganisms are properly broken under the proper and mild ultrasonic condition, and the genomic DNA is prevented from being broken and degraded. As the specific temperature range for incubating the lysozyme and the sample and the specific temperature range for incubating the RNase A and the sample are set, the splitting decomposition effect of the lysozyme can be improved, and it can be avoided that the genomic DNA is degraded by DNA enzymes in tissues and cells. Operation is easy, the extracted genomic DNA is complete and low in degradation rate, and the fish enteric microorganism structure can be truly reflected by the types of the microorganisms identified according to the extracted DNA.

The present invention relates to the technical field of anticancer drugs, and more specifically to a ruthenium complex anticancer drug and a preparation method thereof. The ruthenium complex anticancer drug is activated by redox, inserted into tumor cell DNA, and at the same time coordinates the DNA to undergo cross-linking reaction, prevent DNA replication, and cause DNA breakage. Due to the introduction of planar structure ligand 1,8‑naphthalene dicarboxylic anhydride, the drug’s ability to bind to DNA has been enhanced by 14.17 times, which greatly improves the stability and anticancer properties of anticancer drugs. tumor activity.