417 results about "Chromatin" patented technology

The present invention provides compositions and methods for targeted cleavage of cellular chromatin in a region of interest and / or homologous recombination at a predetermined site in cells. Compositions include fusion polypeptides comprising a TAL effector binding domain and a cleavage domain. The cleavage domain can be from any endonuclease. In certain embodiments, the endonuclease is a Type IIS restriction endonuclease. In further embodiments, the Type IIS restriction endonuclease is FokI.

The invention provides novel methods and materials for genetic and genomic analysis using single or multiplex isolation of protein-associated nucleic acids, including transposase-assisted chromatin immunoprecipitation (TAM-ChIP) and antibody-oligonucleotide proximity ligation. These methods comprise tagging and isolating chromatin or other protein-associated nucleic acids and using antibody-oligonucleotide complexes that recognize the proteins associated with such nucleic acids.

Long non-coding RNAs (lincRNAs), a relatively recently recognized class of widely transcribed genes, are thought to affect chromatin state and epigenetic regulation, but their mechanisms of action and potential roles in human disease are poorly understood. The present invention shows that long non-coding RNAs in the human HOX loci are systematically dysregulated during breast cancer progression, and that expression levels of the lincRNA termed HOTAIR can predict cancer metastasis. Elevated levels of HOTAIR can lead to altered patterns of Polycomb binding to the genome. These findings indicate that lincRNAs have active roles in modulating the cancer epigenome and may be important targets for cancer diagnosis and therapy.

Provided in the present invention is a method for constructing a high-resolution single cell Hi-C library with a large amount of information, comprising the following steps: Step B: obtain a small amount of fixed chromatin; Step C: digest the fixed chromatin in Step B to obtain fragments of the fixed chromatin; Step D: reconnect the fragments of the fixed chromatin in Step C directly to obtain reconnected fragments of the fixed chromatin; Step E: de-fix the reconnected fragments of the fixed chromatin in Step D to release DNA fragments; Step F: amplify the released DNA fragments in Step E to obtain amplified products; and Step H: construct a sequencing DNA library by using the amplified products as the DNA fragments to be sequenced.

A CRISPR / Cas9 system has an ultrahigh parallel capacity. In order to meet the requirement for expressing a plurality of sgRNA in some cases, the invention provides a quick assembling method for a plurality of parallel expressed sgRNA. The invention utilizes grading Golden Gate reaction to develop a multi-turn amplifying method based on polymerase chain reaction and independent of a carrier, so as to realize the quick connecting assembling for 20 sgRNA within one week. The method for serially assembling a plurality of sgRNA developed by the invention has the advantages of time-saving and labor-saving effect, flexibility, high efficiency and multifunction. The method can be used for quickly assembling 2-20 sgRNA in different quantity onto one carrier. A plurality of parallel expressed sgRNA are utilized to target to a section of DNA, so that the functions of marking and tracking unrepeated sequence chromatin locus in living cells, cooperatively activating or restraining a single gene, simultaneously editing a plurality of genes and simultaneously up-regulating and down-regulating the genes can be achieved. The method can be widely used for editing the gene and understanding the organization structure and dynamic change of chromatin.

A non-invasive, risk-free method of prenatal diagnosis is provided. According to the method of the present invention transcervical specimens are subjected to trophoblast-specific immuno-staining followed by FISH, PRINS, Q-FISH and / or MCB analyses and / or other DNA-based genetic analysis in order to determine fetal gender and / or identify chromosomal and / or DNA abnormalities in a fetus. Also provided is a method of in situ chromosomal, DNA and / or RNA analysis of a prestained specimen by incubating the prestained specimen in ammonium hydroxide. Also provided is a method of identifying embryonic cells according to a nucleus / cytoplasm ratio of at least 1:1 and the presence of at least variably condensed chromatin.

The invention concerns the deregulation of the chromatin-regulator genes which have a SET domain, such deregulation being of importance in certain cancer conditions. These genes can be used in the diagnosis and therapy of such conditions.

The invention discloses a construction method for a next-generation sequencing library based on a single-stranded connector and application thereof. The method comprises the following steps: (1) carrying out denaturation on a double-stranded DNA (Deoxyribonucleic Acid) or RNA (Ribonucleic Acid) / DNA heterozygote segment to generate single-stranded DNA; (2) connecting one single-stranded connector with a 3' end of the single-stranded DNA; (3) extending the single-stranded DNA connected with the single-stranded connector by utilizing DNA polymerase to generate double-stranded DNA; (4) connectingthe other end of the double-stranded DNA with a T connector or a Tn5 label connector; (5) carrying out PCR (Polymerase Chain Reaction) amplification on the double-stranded DNA with two ends connectedwith the connectors to form a DNA library capable of sequencing of a next-generation sequencing technology. The method disclosed by the invention can be used for constructing the next-generation sequencing library and determining a DNA sequence and also can be used for identifying a chromatin open region, carrying out gene expression detection, carrying out trace nucleic acid amplification and thelike; the method is a novel method which belongs to the field of nucleic acid detection and analysis and has various functions and wide application value.

The present invention provides novel methods for identifying and monitoring epigenetic modifications, such as imprinted genes, using microarray based technology. Specifically, the invention detects imprinted genes by the presence of overlapping closed and open chromatin markers. The invention also discloses a method for detecting the loss of imprinting on a genome-wide scale, which is indicative of a variety of medical conditions. Diagnostic assays and chromatin structure markers for identifying gene imprinting and loss thereof are also disclosed.

Provided herein are methods and devices for single object detection. The methods and devices can be used to identify a plurality epigenetic markers on a genetic material, or a chromatin, encompassing fragments thereof. The invention provides for the characterization of the genetic material flowing through a channel in a continuous body of fluid based on detection of one or more properties of the genetic material. The methods and systems provided herein allow genome-wide, high-throughput epigenetic analysis and overcome a variety of limitations common to bulk analysis techniques.

A nuclear transfer embryo is formed by destabilizing microtubules of an oocyte, whereby essentially all endogenous chromatin collects at a second polar body during meiosis of an oocyte. The oocyte is fused with the nucleus of a donor somatic cell of the same species of said oocyte prior to cessation of extrusion of the second polar body from the oocyte, thereby forming the nuclear transfer embryo. In one embodiment, the nuclear transfer embryo is employed to impregnate an animal, such as a mammal. In another embodiment, the donor nucleus is transgenic.