32 results about "Beta-thalassaemia" patented technology

The invention provides gRNA for knocking out BCL11A genes or BCL11A gene enhancers, a gRNA composition, an expression vector, CRISPR-Cas9 RNP, a CRISPR-Cas9 RNP composition, a CRISPR-Cas9 system, an electrorotation method, a reagent kit and applications thereof. A CRISPR-Cas9 technique is adopted to perform targeted cutting on hemopoietic stem cells, so that the BCL11A gene expression amount is reduced, and the hemochrome expression quantity of fetuses is high. The technique is hopeful to become a new means for treating thalassaemia and sickle cell anemia. The CRISPR-Cas9 technique is adoptedto realize mutation of BCL11A genes. The design is simple, the use is convenient, the cost is low, and the efficiency is high.

The invention discloses primer pairs and probes for detecting thalassemia genes. The primer pairs and probes include primer pairs and probes of beta-thalassemia CD41 / 42 and alpha-thalassemia-SEA; theprimer pair of the beta-thalassemia CD41 / 42 is a sequence 1 and a sequence 2, and the probes are a sequence 3 and a sequence 4; and the primer pair of thealpha-thalassemia-SEA is a sequence 5 and a sequence 6 or a sequence 5 and a sequence 7, and the probes are a sequence 8 and a sequence 9. The invention further provides a use method of the primer pairs and probes for detecting the thalassemia genes. The use method comprises the following steps of S1, collecting a sample and extracting DNA; S2, carrying out an amplification reaction by utilizing one kind of primer pair and probes; S3, detecting the fluorescence intensity and determining the genotype; and S4, repeating the process by utilizing the other kind of primer pair and probes. The method has the beneficial effects that the fluorescence detection accuracy is enhanced through the designed primer pairs and probes and the two kinds of primer pairs of the same type; and rapid, accurate and harmless detection is realized by extracting free DNA of plasma and blastocyst culture media of pregnant women.

The invention discloses an alpha-thalassemia related gene detection kit. The invention provides an alpha-thalassemia detection primer group for detecting alpha globin fusion gene, particularly primers with nucleotide sequences as shown in SEQ ID NO: 2-3, and a detection kit constructed based on the alpha-thalassemia detection primer group; and the operation is simple, the cost is low, the specificity is high, the result interpretation is visual, the popularization is easy, and the alpha-thalassemia detection primer group is suitable for being used in various occasions. Moreover, the primer group also has a very good multi-system general advantage, and can be matched with other detection primers for multiple PCR detection, for example, primers with nucleotide sequences as shown in SEQ ID NO: 4-5 or primers with nucleotide sequences as shown in SEQ ID NO: 6-7 can be used for detecting various types of alpha-thalassemia at one time. The technical scheme provided by the invention is of great significance to screening, genetic counseling and prenatal diagnosis of crowds suffering from thalassemia.

The present invention relates to methods and pharmaceutical compositions for the treatment of beta-thalassemias. In particular, the present invention relates to an XPO1 inhibitor for use in a method for treating beta-thalassemia in a subject in need thereof.

The invention discloses an α-thalassemia screening kit and its application in prenatal screening, belonging to the technical field of α-thalassemia prenatal screening. The α-thalassemia screening kit includes: (1) Negative control sample; (2) Positive control sample; (3) PCRMasterMIX; (4) PCR primers. Its application uses designed primers to amplify fetal DNA in the peripheral blood of pregnant women to achieve prenatal screening for α-thalassemia. The present invention collects the peripheral blood of pregnant women for prenatal screening of α-thalassemia, without puncturing the amniotic cavity and inserting villi tissue, without any trauma to the fetus, and is safe and reliable; the accuracy rate can reach 99.99%, filling the non-invasiveness of α-thalassemia The technical gap in prenatal screening reduces the birth rate of sick children.

The invention provides an application of thyroid hormone and an analogue thereof in preparation of a drug for treating alpha-thalassemia. The thyroid hormone and the analogue can be specifically applied in regulation of expression of zeta-globin genes, in the differentiation process of K562 cells, the thyroid hormone analogue can be specifically and significantly up-regulate the expression of thezeta-globin gene (HBZ) by up to 50 times, and in model animal zebrafish embryos, after treatment is performed by using the thyroid hormone and the analogue, the expression of the zeta-globin gene (hbae5) can also be specifically up-regulated by up to 30-70 times; therefore, the thyroid hormone and the analogue thereof can be used to specifically activate the expression of the zeta-globin genes, that is, to reactivate the silent zeta-globin genes in patients with the alpha-thalassemia to inhibit destruction of red blood cells, the thyroid hormone and the analogue realizes a method for preparingthe drug for treating the alpha-thalassemia, the economical, safe and effective method is provided for treatment of the alpha-thalassemia, and the method can be widely promoted and used.

The invention discloses a method and product for hematopoietic stem cell HBB gene repair. The method utilizes CRISPR-Cas9 gene editing technology to target knockout of codon 41 / 42(-TCTT) in β-thalassemia (thalassemia) ) technology, by designing and synthesizing sgRNA that can recognize and guide Cas9 protein to the target sequence of the target gene, mix it with Cas9 protein and electrotransduce it into β-thalassaemia codon 41 / 42 (‑TCTT) hematopoietic stem cells, and introduce homologous The recombinant donor efficiently restores the normal coding function of the amino acid at the mutation site and restores the normal expression of the β-globin gene. The present invention utilizes the existing gene editing technology to edit transfusion-dependent β-thalassemia codon 41 / 42 (-TCTT), which has high repair efficiency, and the repaired patient's hematopoietic stem cells can rebuild the patient's blood system and treat thalassemia after autologous transplantation .

The present invention provides a detection primer, method, chip and application of α and / or β-thalassemia point mutations and deletion mutations based on whole gene capture sequencing. The present invention enriches all related genes involved in α and β thalassemia by designing capture probes, detects mutation information such as all SNPs and indels in the full-length sequences of each gene, and simultaneously increases The upstream and downstream regions of each coding gene are used as a reference to detect structural variations such as SNVs and CNVs. Compared with the current detection techniques of various hotspot mutation sites, the present invention can not only detect the information of hotspot mutations, but also detect some rare mutations and undiscovered new mutation types, and realize the specific detection and analysis of the full-length sequence of the target gene. Full coverage of types, greatly making up for the missed detection of low-frequency mutations and rare mutations by conventional detection methods.

The invention relates to the technical field of biology and medicament, and particularly relates to a gene detection kit for Hong Kong alpha-thalassemia. According to the invention, the gene sequence of HK alpha alpha fusion gene is further confirmed and compared with alpha-globin sequence for analysis, and primer design is carried out in a conserved sequence area of the fusion gene. The technical scheme of the invention is to provide a gene detection kit for Hong Kong alpha-thalassemia, comprising a PCR (polymerase chain reaction) solution, wherein the PCR solution contains primer HK alpha alpha-F and HK alpha alpha-R, the primer is designed aiming at the conserved sequence area of the HK alpha alpha fusion gene, and the nucleotide sequence of the conserved sequence of the HK alpha alpha fusion gene is shown in SEQ ID NO:1. The kit disclosed by the invention can be directly used for kits for HK alpha alpha detection and can specifically, quickly and stably diagnose HK alpha alpha gene type.

The invention provides a capture probe composition for three subtypes of thalassemia, an application method and an application device. Wherein, the preparation method of the capture probe composition of the three subtypes of thalassemia includes: separating the full-length regions of the genes related to the three subtypes of thalassemia into different small fragments according to preset lengths; The fragments are compared to the human reference genome to obtain the first single alignment region set and multiple alignment region sets; move the multi-alignment region set forward or backward on the human reference genome by a length of kbp, k≤100, and move The final new region is re-aligned with the human reference genome to obtain a second single alignment region set; probe design is performed from the first single alignment region set and the second single alignment region set to obtain a capture probe composition. The capture probe composition obtained by the method can not only mark the whole region but also can distinguish three different thalassemia types, so that the detection is more accurate.

The invention relates to in vitro molecular diagnosis technology, in particular to a primer combination and a kit for detecting deletion type α-thalassemia. The primer combination can stably detect -α 3.7 and-(α) 20.5 The two deletion types overcome the problems of high GC content in the α-globin gene region and unstable amplification system; at the same time, the annealing temperature of each primer is within the same temperature range, which avoids non-specific amplification caused by different annealing temperatures. The phenomenon that the amplification efficiency varies. In addition, the primers have good compatibility and no dimer is produced. The kit overcomes the influence between primers, and can detect 7 genotypes of thalassemia in a single tube and multiplex. The need for rapid and convenient clinical detection of thalassemia creates conditions for a more comprehensive screening of thalassemia, and provides a scientific basis for premarital and prenatal examinations and the diagnosis of thalassemia in fetuses during pregnancy.