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Myeloid cell promoter and constructs containing sameRelated Patent Categories: Chemistry: Molecular Biology And Microbiology, Process Of Mutation, Cell Fusion, Or Genetic Modification, Introduction Of A Polynucleotide Molecule Into Or Rearrangement Of Nucleic Acid Within An Animal CellMyeloid cell promoter and constructs containing same description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20050266562, Myeloid cell promoter and constructs containing same. Brief Patent Description - Full Patent Description - Patent Application Claims
TECHNICAL FIELD [0001] The present invention relates to a novel promoter isolated from a myeloid cell, as well as cis-acting elements from the myeloid cell, and to novel constructs containing the same. BACKGROUND OF THE INVENTION [0002] The expression of foreign genes in various cell types has become a commonplace occurrence in the field of biotechnology. The expression of the foreign, or heterologous, gene is performed by the transcription of the genetic information in the gene from DNA to RNA. The RNA is then translated into the protein for which the gene encodes. [0003] In transcription, the RNA is synthesized using the DNA of the gene for a template. The RNA is synthesized by a reaction catalyzed by RNA polymerases. The RNA polymerase binds to a particular molecular site on the DNA to initiate the transcription process. This site to which the RNA polymerase binds is known as the promoter. [0004] Therefore, in order to express heterologous genes in foreign cells, the genes must be under the control of a promoter. Although numerous promoters for expressing foreign genes have been taught in the literature, these promoters have generally failed to function when used to express genes within myeloid cells. Therefore, a need exists for promoters which may be used to express genes within such cells. [0005] Integrins are a large family of cell surface glycoproteins that are heterodimers comprised of .alpha. and .beta. chain subunits. The promoters of integrins are of special interest as these promoters may be used to express genes in a myeloid-specific manner. This is especially useful, as typical promoters which are used in genetic research, such as retroviral promoters, become repressed after being introduced into myeloid cells. [0006] Genes linked to myeloid specific promoters may be used in a wide variety of applications. These applications include use in the development of cancer vaccines, as well as being used for screening compounds for their effect on myeloid cell specific gene expression. [0007] A need therefore remains for the identification and isolation of myeloid cell specific promoters. SUMMARY OF THE INVENTION [0008] The present invention relates to myeloid cell specific gene expression. The invention includes myeloid cell specific gene expression under the control of a myeloid cell specific promoter, preferably the CD11d promoter. [0009] A preferred myeloid cell specific promoter is all or a functional portion of isolated or recombinant SEQ ID NO: 1, such that the sequence is sufficient to direct myeloid cell specific expression of a gene. More preferably, the myeloid cell specific promoter of the present invention is the -946 to +74 region that is upstream (5') of the CD11d gene, as identified by SEQ ID NO: 1. In particular, the present invention relates to portions of this sequence that are sufficient to direct myeloid cell specific expression of a heterologous gene, and also includes modifications of this sequence that retain sufficient activity to direct myeloid cell specific expression of a heterologous gene. More preferably, the invention relates to the region from -173 to +74 that is upstream (5') of the CD11d gene. The invention further includes all or a functional portion of isolated or recombinant SEQ ID NO: 1, wherein the sequence comprises one or more modifications, such that the modified sequence retains sufficient activity to direct myeloid cell specific expression of a gene. [0010] The invention further relates to a cis-acting element that influences the activity of the myeloid cell specific promoter. This influence may be either to increase or decrease the activity of the myeloid cell specific promoter. The cis-acting element comprises a portion of SEQ ID NO: 1 that is sufficient to influence the activity of the myeloid cell specific promoter. In particular, the present invention relates to portions of this sequence that are sufficient to function as a cis-acting element that influences the activity of the myeloid cell specific promoter, and also includes modifications of this element that retain sufficient activity to influence the activity of the myeloid cell specific promoter. [0011] A preferred cis-acting element is the -173 to +74 region that is upstream (5') of the CD11d gene as shown in SEQ ID NO:1. Also preferred is the -72 to -40 region, more specifically, the -63 to -40 region, that is upstream (5') of the CD11d gene. In particular, the invention is directed to cis-acting elements that positively influence the CD11d promoter, increasing the activity of the promoter. [0012] Cis-acting elements that negatively influence the CD11d promoter are also incorporated as part of the present invention. Particularly, the present invention relates to the -591 to -378 region that is upstream (5') of the CD11d gene as shown in SEQ ID NO:1. This cis-acting element contains a cell-specific silencer element. [0013] The invention also relates to the approximately 12 kb sequence that is upstream (5') of the CD11d gene as shown in SEQ ID NO:2. This sequence further comprises additional cis-acting elements which influence the activity of the myeloid cell promoter of the present invention. [0014] Additionally, the invention includes constructs that contain DNA sequences sufficient to direct myeloid cell specific expression of a gene. These constructs comprise both the promoter of the present invention and a heterologous gene, whereby the expression of the heterologous gene is under the control of the myeloid cell specific promoter. The invention is also directed to constructs that contain one or more cis-acting elements of the present invention together with the promoter of the present invention, both linked to a heterologous gene, whereby the expression of the heterologous gene is under the control of the myeloid cell specific promoter of the present invention, and the promoter is influenced by the cis-acting element(s) of the present invention. [0015] The invention also includes cells that have been transfected with a construct of the present invention. [0016] In addition, the present invention relates to methods of producing a selected heterologous gene product in a myeloid cell. These methods include introducing into the myeloid cell a heterologous gene under the control of a myeloid cell specific promoter, or a cis-acting element that influences the activity of the myeloid cell specific promoter, or both. The invention also includes cells produced by the methods described herein. [0017] The present invention also relates to a method for identifying factors that can regulate myeloid cell specific transcription. [0018] Further, the invention relates to a method of expressing a selected heterologous gene product in myeloid cells of an individual, i.e., gene therapy. According to this embodiment, cells produced by the methods described herein are introduced into an individual, wherein they express a heterologous gene under transcriptional control of a myeloid cell specific promoter or a cis-acting element that influences the activity of the cell specific myeloid promoter, or both. [0019] The present invention provides a means of insuring that a selected product, such as a diagnostic, therapeutic or prophylactic substance, is expressed from a specific cell type, in vivo. Therefore, the present invention is useful, for example, for gene therapy or to drive the expression of antiviral agents, such as anti-HIV constructs. The present invention also provides novel promoters which may used to produce cancer vaccines. The present method is also useful in research, for example, to test the effect of the specific expression of heterologous genes, such as oncogenes, in specific cell types. Cells of the present invention are also useful for screening compounds for their effect on myeloid cell specific gene expression. [0020] Further objects, features and advantages of the invention will be apparent from the following detailed description when taken in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS Continue reading about Myeloid cell promoter and constructs containing same... 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