87 results about "Globin genes" patented technology

This invention relates to test kit and test paper with oligonucleotide probes for diagnosing alpha-thalassemia. The test paper comprises a base, and specific oligonucleotide probes immobilized on the base. The oligonucleotide probes comprise: specific oligonucleotide probes for detecting mutation of alpha-globin gene, specific oligonucleotide probes for detecting deletion of alpha-globin gene, and base sequence complementary with the above sequences. The test kit comprises specific primer for amplifying alpha-globin gene or transcript, and specific oligonucleotide probes for detecting mutation / deletion of alpha-globin gene. The test kit and the test paper have such advantages as low cost, rapid detection, and stable results. The test kit and the test paper can be used in detection of mutation / deletion of alpha-globin gene, and prenatal diagnosis of severe thalassemia (alphaTalpha) caused by point mutation, and do not have error diagnosis.

A multiple PCR detection method for the depletion-type human alpha-globin gene features that 4 pairs of 7 primers P1, P2, P3, p4, P5, P6 and P7 are used to form quadriplex PCR reaction. The design scheme of said primers and the explanation of result are also disclosed. It can be used for screening and diagnosing three types of alpha-Mediterranean anemia.

The invention belongs to the field of biochemistry, and provides a fluorescent quantitative polymerase chain reaction (PCR) kit for detecting alpha-globin gene deletion. The kit consists of a pair of specific amplified xi globin primers, a pair of specific amplified alpha1 globin primers, a pair of specific amplified alpha2 globin primers, a pair of specific amplified beta-Actin gene primers, a fluorescent probe for specifically detecting xi globin, a fluorescent probe for specifically detecting alpha1 globin, a fluorescent probe for specifically detecting alpha2 globin, a fluorescent probe for specifically detecting beta-Actin genes, a DNA polymerase and the like. The kit has good sensitivity and accuracy for detecting depletion alpha-thalassemia, has good repeatability and stability, and is totally suitable for clinical detection of alpha-thalassemia.

The invention provides an alpha-globin gene mutation detection kit as well as a preparation method and a use thereof, and relates to protein gene mutation detection. The alpha-globin gene mutation detection kit is provided with a box, an amplification reagent bottle and a control reagent bottle; the amplification reagent bottle and the control reagent bottle are arranged in the box. The preparation method comprises the following steps: 1) preparing an amplification reagent which comprises an HBA PCR mixed solution and an HBA enzyme mixed solution; 2) preparing a control reagent which comprises HBA standard control and HBA negative control; and 3) putting the amplification reagent prepared in the step 1) and the control reagent prepared in the step 2) in the box, thereby obtaining the alpha-globin gene mutation detection kit. The alpha-globin gene mutation detection kit can be applied to detecting the alpha-globin gene mutation. The alpha-globin gene mutation detection kit is convenient and fast, capable of detecting a plurality of sites by use of one single reaction, short in time consumption, high in detection flux, high in detection specificity, and easy in result interpretation.

The invention discloses a human beta globin gene containing a human beta globin gene promoter, a recombinant adeno-associated virus vector containing the human beta globin gene, and a method for preparing the recombinant vector. The recombinant adeno-associated virus vector inserts an HS2 segment, an HS3 segment and an HS4 segment of a human beta globin gene cluster enhancer core sequence and a human beta globin gene sequence containing the human beta globin gene promoter into repeated sequence ITRs on the reverse terminal of an adeno-associated virus. The recombinant vector has high transfection efficiency; and mediated exogenous genes can be expressed in vivo for a long time, have good safety and can be used for gene therapy of beta Mediterranean anemia.

The invention provides a novel HBB overexpression vector and a design method and application thereof. The HBB overexpression vector comprises an HBB expression module; the HBB expression module comprises a DNase I core high-sensitivity site, a promoter, an HBB expression frame and a downstream high-sensitivity site which are arranged in series; the DNase I high-sensitivity site comprises HS4, HS3,HS2, 3'E and the like which are expressed in series. The total length of the HBB expression module is smaller than 4 kb. By simplifying and optimizing a cis-acting element and the HBB expression frame, not only is the length of the HBB expression module obviously shortened, but also the transcriptive activation strength of the HBB expression module is improved, and efficient, stable and specificactivation of the HBB globin gene is realized.

The invention discloses a nested asymmetric PCR reagent kit for detecting alpha 2 globin gene point mutation. The reagent kit comprises a nested asymmetric PCR amplification primer, a fluorescent probe, a quenching probe, an LADNA polymerase and the like. The alpha-thalassemia point mutation genotype of a detected sample is detected by virtue of specific amplification of five point mutation destination fragments WS, QS, CS, CD30 and CD31 of the human alpha 2 globin gene and molecular hybridization and unwinding analysis of a specific fluorescent probe. The reagent kit has good sensitivity and accuracy to wild type and mutation type of alpha-thalassemia five point mutation positions WS, QS, CS, CD30 and CD31, has good repeatability and stability, and is totally suitable for clinical detection of alpha-thalassemia point mutation.

The invention provides a method and a system for detecting alpha-globin gene copy number in a nucleic acid sample. The method comprises the following steps of: amplifying the nucleic sample to obtain an amplification product; establishing a sequencing library for the amplification product; sequencing the sequencing library to obtain a sequencing result, wherein the sequencing result is composed of multiple sequencing data; determining the sequencing data from alpha-globin gene in the sequencing result; and determining the copy number of the alpha-globin gene in the nucleic acid sample based on the number of the sequencing data of the alpha-globin gene. The method provided by the invention can effectively determine the copy number of the alpha-globin gene in the nucleic acid sample.

The presently disclosed subject matter provides for expression cassettes that allow for expression of a globin gene or a functional portion thereof, vectors comprising thereof, and cells transduced with such expression cassettes and vectors. The presently disclosed subject matter further provides methods for treating a hemoglobinopathy in a subject comprising administering an effective amount of such transduced cells to the subject.

Recombinant lentiviral vectors having a region encoding a functional β-globin gene; and large portions of the β-globin locus control regions which include DNase I hypersensitive sites HS2, HS3 and HS4 provides expression of β-globin when introduced into a mammal, for example a human, in vivo. Optionally, the vector further includes a region encoding a dihydrofolate reductase. The vector may be used in treatment of hemoglobinopathies, including β-thalessemia and sickle-cell disease. For example, hematopoietic progenitor or stem cells may be transformed ex vivo and then restored to the patient. Selection processes may be used to increase the percentage of transformed cells in the returned population. For example, a selection marker which makes transformed cells more drug resistant than un-transformed cells allows selection by treatment of the cells with the corresponding drug.

The invention discloses a kit for detecting common deletion form alpha-thalassemia and a using method of the kit. The kit comprises a pair of primers capable of performing simultaneous amplification of a signature sequence A1 in an alpha 1 section and a signature sequence A2 in an alpha 2 section in an alpha-globin gene cluster, a pair of primers capable of performing amplification of a --<SEA> genetype in the alpha-globin gene cluster and a pair of primers capable of performing amplification of a --<THAI> genetype in the alpha-globin gene cluster as well as a fluorescence probe for specific detection of the signature sequence A1 in the alpha 1 section, a fluorescence probe for specific detection of the signature sequence A2 in the alpha 2 section, a fluorescence probe for specific detection of a --<SEA> genetype amplicon and a fluorescence probe for specific detection of a --<THAI> genetype amplicon. As for the alpha-thalassemia globin gene deletion detection, the kit provided by the invention has high sensitivity, stability, accuracy and specificity.

The orphan nuclear receptors TR2 and TR4 together constitute the DNA binding core of the 540 kDa DRED complex, a putative repressor of the human embryonic ε- and fetal γ-globin genes. Here the functional consequences of TR2 and TR4 germ line loss of function were examined, transgenic gain of function and dominant negative gain of function on human and murine β-type globin gene expression throughout development. ε-globin transcription responded in a manner consistent with the hypothesis that TR2 / TR4 is a constitutive erythroid ε-globin repressor. In contrast, parallel experiments show that TR2 / TR4 is a definitive stage-selective γ-globin repressor. This developmental stage-specific, gene-selective repression of the ε- and γ-globin genes by TR2 / TR4 establishes, when considered in concert with the competition hypothesis, a coherent molecular rationale for hemoglobin switching (temporally specific, sequential activation of all the β-type globin genes) during vertebrate development.

The invention is directed to a system comprising a lentivirus vector particle which encodes at least one guide RNA sequence that is complementary to a first DNA sequence in a host cell genome, a Cas9 protein, and optionally a donor nucleic acid molecule comprising a second DNA sequence. The invention also is directed to a method of altering a DNA sequence in a host cell using such a system, where the host cell can be in a human and the altered DNA can be of the human β-globin gene. The invention also is directed to a fusion protein comprising a Cas9 protein and a cyclophilin A (CypA) protein. The invention also is directed to sequences of vectors that can be used in the system and method.

The present invention relates to vectors comprising an alpha -globin locus control region ( alpha LCR) and a gene encoding an erythroid protein. In particular embodiments, a retroviral vector comprising an alpha LCR and a globin gene may be used to treat globin-based genetic disorders, including sickle cell anemia and beta -thalassemia.

The invention discloses a probe for detecting common genetic diseases, a gene chip, a preparation method and application. The preparation method comprises the following steps of according to genes ofthalassemia, phenylketonuria, spinal muscular atrophy, hepatolenticular degeneration, pseudomuscular hypertrophy, glycogen storage disease Type II, methylmalonic academia, methylmalonic aciduria-accompanied type cysteine peptiduria Type cb1C, and oculocutaneous albinism Type 1, constructing a target capturing area, and designing a capturing probe, wherein the target capturing area comprises all globin genes, important regulating areas, deleted mutation breakpoints and SNP (single nucleotide polymorphism) sites of alpha and beta genes, and SMN1, PAH, ATP7B, DMD, GAA, MUT, MMACHC and TYR genes.The probe for the gene mutation of common genetic diseases can be designed according to the related genes of nine types of genetic diseases, and the prepared probe can be used for detecting the nine types of genetic diseases, and be suitable for large-scale population diagnosis and screening.

The invention provides a human papillomavirus gene chip, a preparation method and application thereof, an assay kit and application thereof. The gene chip comprises a solid-phase vector and an oligonucleotide probe fixed on the solid-phase vector, wherein the oligonucleotide probe contains a DNA fragment or a complementary DNA fragment thereof selected from L1 genes of the human papillomavirus or human beta globin gene. The invention also provides a preparation method for the gene chip and an assay kit containing the gene chip. The gene chip and the kit have the characteristics of simple and convenient operation, high flux, high accuracy, strong repeatability and the like, can achieve the aim of detecting 25 different types of human papillomavirus, and can be applied to the clinical test, epidemiology analysis and the like of medical and health departments.

The invention discloses a probe and a gene chip for thalassemia detection, a preparation method and applications. The preparation method of the probe for the thalassemia detection includes the following steps: constructing a capture interval library based on different thalassemia genes, modified genes and mutations; and designing a capture probe according to the capture interval library, wherein the capture interval library includes all globin genes and important regulatory region full-length fragments of an alpha gene cluster and a beta gene cluster, recorded breakpoint mutation sites, SNP sites selected from deletion mutations, and SNP sites on thalassemia modified genes. The preparation method of the invention is to design the probe aiming at all gene clusters and related genes of thalassemia, and so the probe can cover all thalassemia gene mutations. The preparation method is combined with Tn5 library construction and high-throughput sequencing, and so all the mutations can be covered, and the method has the advantages of simple operation, high throughput and low cost, and is suitable for large-scale population diagnosis and screening of thalassemia and research and predictionof new unknown gene mutations.

The invention discloses a method for rapidly detecting the copy number variation of alpha-globin gene cluster, which comprises the following steps: detecting one pair of universal primers, three kinds of TaqMan probes and three pairs of tailing primers, and detecting a sample: adding the three pairs of tailing primers by taking a gene to be detected and a standard gene as templates, carrying out the first round of nested fluorescent quantitative PCR (polymerase chain reaction) to obtain a first round of products of PCR, adding the universal primers and three kinds of TaqMan probes by taking the first round of products of PCR as templates, carrying out the second round of fluorescent quantitative PCR, collecting fluorescent signals after finishing each cycle of reaction, and quantifying the copy numbers of both the alpha1-globin gene and the alpha2-globin gene according to the collected fluorescent signals. Due to the adoption of the method, the reaction efficiency for amplification of a plurality of target segments in the same reaction pipe is consistent, and enotypes of the alpha-globin gen can be distinguished accurately. The method is easy to operate and has high throughput and requires a two-step PCR cycle reaction of 2h or shorter.

The invention relates to a thalassemia-induced multipotent stem cell as well as a preparation method and application thereof. The multipotent stem cell is derived from a body cell of a thalassemia patient; and a genome of the thalassemia-induced multipotent stem cell contains a normal beta globin gene. The thalassemia-induced multipotent stem cell disclosed by the invention can be differentiated into hemopoietic stem cells under proper conditions; and the hemopoietic stem cells can express the normal beta globin gene.

A method is described for regulating gene expression related to iron metabolism to ameliorate diseases that include sickle cell disease, cancers, neurodegenerative diseases, Friedreich's ataxia and other neuromuscular disorders, and atherosclerosis. This approach is illustrated by recent findings that show that ferritin-H, an iron-binding protein that is present in cell nuclei, can repress the human .beta.-globin gene, the gene that is mutated in sickle cell disease. Increased expression of ferritin-H or a related ferritin-family peptide, given to effected cells either as the peptide itself (or a part thereof), as an expression clone of the ferritin-H-subfamily gene, or via a gene regulator that increases expression of the ferritin-H-subfamily gene itself, prevents or ameliorates expression of the disease state in disorders where increased availability of iron is implicated in the etiology of the disease, including those named above.

The present invention provides a globin gene dual-expression lentivirus vector and an application thereof. The lentiviral vector comprises optimized multiple globin genes connected by P2A-T2A connecting peptides. The efficient expression of globin is realized in protein level, the length of the vector is substantially reduced, the lentivirus packaging efficiency is remarkably improved, the complexity and quality control requirements of a production process are lowered, the problems of low efficiency and high energy consumption of the globin lentivirus vector are solved fundamentally, and an innovative scheme is provided for high-cost-performance large-scale stable production of gene therapeutic drugs.

The invention relates to the technical field of medical detection, in particular to primers for detecting mycoplasma genitalium, a probe for detecting mycoplasma genitalium, a kit for detecting mycoplasma genitalium and a detection method. The primers and the probe comprise a primer pair of nucleotide sequences shown in SEQ ID NO:1-2 as shown in the description and a probe of a nucleotide sequenceshown in SEQ ID NO:3 as shown in the description. The provided primers and probe can specifically bound to a mycoplasma genitalium outer membrane protein B gene and a human beta-globin gene, the detection sensitivity and specificity are significantly improved, the positive rate of low-value samples reaches 100%, and detection results are accurate and reliable so that the primers, probe and kit for detecting mycoplasma genitalium and the detection method can be applied to clinical detection of mycoplasma genitalium.

The invention provides application of a CRISPR (clustered regularly interspaced short palindromic repeats) gene editing technology in treatment of thalassemia. A CRISPR system in the application comprises at least one nuclease and at least one sgRNA, the sgRNA can target an inhibiting element of an HBG (gamma globin gene) promoter; the nuclease can cut the inhibiting element of the HBG promoter, the sgRNA comprises a targeting sequence targeting the inhibiting element of the HBG promoter, and the targeting sequence comprises a sequence as shown in SEQ ID No.1 or a complementary sequence as shown in SEQ ID No.1. Transferring a compound of the nuclease and the sgRNA into a cell by an electrotransformation method can knock out the inhibiting element on the HBG (gamma globin gene) promoter, and the relative content of HBG mRNA in cells is improved, and compared with a traditional lentiviral infection technology, exogenous DNA random integration caused by viruses is avoided, and the prospect of clinical application is improved.