86 results about "Globin genes" patented technology

Provided are compositions and methods for the treatment of hemoglobinopathies such as thalassemias and sickle cell disease. Compositions and methods include one or more endonuclease(s) or endonuclease fusion protein(s), including one or more homing endonuclease(s) and / or homing endonuclease fusion protein(s) and / or CRISPR endonuclease(s) ad / or CRISPR endonuclease fusion protein(s): (a) to disrupt a Bcl11a coding region; (b) to disrupt a Bcl11a gene regulatory region; (c) to modify an adult human β-globin locus; (d) to disrupt a HbP silencing DNA regulatory element or pathway, such as a Bcl11a-regulated HbP silencing region; (e) to mutate one or more γ-globin gene promoter(s) to achieve increased expression of a γ-globin gene; (f) to mutate one or more δ-globin gene promoter(s) to achieve increased expression of a δ-globin gene; and / or (g) to correct one or more β-globin gene mutation(s).

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.

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 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.

Provided are compositions and methods for the treatment of hemoglobinopathies such as thalassemias and sickle cell disease. Compositions and methods include one or more endonuclease(s) or endonuclease fusion protein(s), including one or more homing endonuclease(s) and / or homing endonuclease fusion protein(s) and / or CRISPR endonuclease(s) and / or CRISPR endonuclease fusion protein(s): (a) to disrupt a Bcl11a coding region; (b) to disrupt a Bcl11a gene regulatory region; (c) to modify an adult human β-globin locus; (d) to disrupt a HbF silencing DNA regulatory element or pathway, such as a Bcl11a-regulated HbF silencing region; (e) to mutate one or more γ-globin gene promoter(s) to achieve increased expression of a γ-globin gene; (f) to mutate one or more δ-globin gene promoter(s) to achieve increased expression of a δ-globin gene; and / or (g) to correct one or more β-globin gene mutation(s).

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 untransformed cells allows selection by treatment of the cells with the corresponding drug.

The invention discloses a lentiviral vector suitable for gene therapy of thalassemia and sickle anemia. The system comprises: a, a micro locus control region, which is a micro regulatory element which is screened from a locus control region of beta globin 16kb and does not contain an HS1 region; b, a gene sequence of beta globin, wherein the gene sequence is as shown in SEQ ID NO.3; c, a promoter sequence of an upstream flanking of the gene sequence of beta globin; d, a flanking sequence at the downstream of the gene sequence of beta globin; and e, an insulator sequence from a Foamy virus (Foamy virus). Compared with the prior art, the invention optimizes the structure of the vector, and finds that after the second intron of the Beta globin gene (HBB) is optimized, the expression of the gene can be improved to a certain extent, and the packaging yield of the virus can be greatly improved, so that the clinical use and industrialization of the Beta globin gene become possible.