# US 8,541,386 B2 · Cannabinoid 2 (CB2) receptor gene promoter and unique RNA transcripts in B cells and methods of use > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,541,386 B2 - Document ID: US8541386B2 - Status: Granted - Kind: B2 - Application number: 12/859,744 - Filed: 2010-08-19 - Published: 2011-04-21 - Granted: 2013-09-24 - Earliest claimed: 2009-08-19 - Assignees: University of South Florida - Applicants: Thomas W. Klein; Tracy Sherwood; Liang Nong; Cathy Newton - Inventors: Thomas W. Klein; Tracy Sherwood; Liang Nong; Cathy Newton - Web page: https://patentgrep.com/patent/US8541386B2 ## CPC - C12Q 1/68: involving nucleic acids - A61K 31/7105: Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links - A61P 11/06: Antiasthmatics - A61P 37/00: Drugs for immunological or allergic disorders - C07K 14/70571: {for neuromediators, e.g. serotonin receptor, dopamine receptor} - C12N 15/1138: {against receptors or cell surface proteins} - C12Q 1/6883: for diseases caused by alterations of genetic material - C12N 2310/11: Antisense - C12N 2310/14: interfering nucleic acids [NA] - C12Q 2600/106: Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract Cannabinoid receptor 2 (CB2) is expressed in B lymphocytes and is involved in immune regulation. Mouse splenic B cells express three CB2 transcripts utilizing two different first exons. Human peripheral blood B cells express one CB2 transcript utilizing one first exon. Alignment of sequenced RACE products to either the mouse or human genome reveals that isolated transcripts contain previously unidentified transcriptional start sites (TSS). B cells from mouse and human preferentially express one transcript, exon 1a in mouse and exon 1 in human. Multiple CB2 TSSs are utilized in mouse splenic B cells and one TSS in human peripheral blood B cells. The defining of the receptor gene TSSs in these cells provides materials and methods for therapeutically regulating immune function, including antibody isotype switching, using compounds such as inhibitory nucleic acids that down-regulate expression of the B cell CB2 gene (e.g., RNAi molecules). ## Claims ### Claim 1 A method of reducing the expression of a B cell CB2 receptor gene, comprising contacting the B cell with one or more inhibitory nucleic acid molecules comprising a nucleic acid sequence that targets a sequence (nucleic acid target sequence within or overlapping with the transcription start site associated with the B cell CB2 receptor gene promoter region within the cell, thereby reducing expression of the B cell CB2 receptor gene. ### Claim 2 (depends on 1) The method of claim 1, wherein the one or more inhibitory nucleic acid molecules are selected from among an interfering RNA (RNAi) molecule, an antisense oligonucleotide, a ribozyme, and a construct that causes targeted deletion of the CB2 receptor gene. ### Claim 3 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence comprises at least one region selected from among the transcription start site (tss) for human exon 1, or at least a portion of the 5′ untranslated region (UTR) for human exon 1. ### Claim 4 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence comprises at least one region selected from among the transcription start site (tss) for mouse exon 1a, mouse exon 1b, or mouse exon 2, or at least a portion of the 5′ UTR for mouse exon 1a, mouse exon 1b, or mouse exon 2. ### Claim 5 (depends on 1) The method of claim 1, wherein the one or more inhibitory nucleic acid molecules are contacted with the cell in vivo, and wherein said contacting comprises administering the one or more inhibitory nucleic acid molecules to a subject. ### Claim 6 (depends on 5) The method of claim 5, wherein said administering results in regulation of an immunoglobulin E (IgE)-mediated allergic response in the subject. ### Claim 7 (depends on 1) The method of claim 1, wherein the cell is a human or mouse cell. ### Claim 8 (depends on 1) The method of claim 1, wherein the one or more inhibitory nucleic acid molecules are contacted with the cell in vitro. ### Claim 9 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence comprises a region surrounding and including the 5′ end of human exon 1. ### Claim 10 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence comprises a region surrounding and including the 5′ end of mouse exon 1a, the 5′ end of mouse exon 1b, or the 5′ end of mouse exon 2. ### Claim 11 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence comprises a region surrounding and including the 5′ end of mouse exon 1b. ### Claim 12 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence comprises a region surrounding and including the 5′ end of exon 2. ### Claim 13 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence is a sequence within or overlapping with nucleotides 83-117 of SEQ ID NO:121. ### Claim 14 (depends on 1) The method of claim 1, wherein the nucleic acid sequence of the one or more inhibitory nucleic acid molecules is complementary to a sequence within or overlapping with nucleotides 83-117 of SEQ ID NO:121. ### Claim 15 (depends on 1) The method of claim 1, wherein the nucleic acid sequence of the one or more inhibitory nucleic acid molecules is sufficiently complementary to a sequence within, or overlapping with, nucleotides 83-117 of SEQ ID NO:121 to direct target-specific RNA interference (RNAi). ### Claim 16 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence is a sequence within or overlapping with: nucleotides 1-14 of SEQ ID NO:118, or nucleotides 1-294 of SEQ ID NO:119, or nucleotides 1-172 of SEQ ID NO:120. ### Claim 17 (depends on 1) The method of claim 1, wherein the nucleic acid sequence of the one or more inhibitory nucleic acid molecules is complementary to a sequence within or overlapping with: nucleotides 1-14 of SEQ ID NO:118, or nucleotides 1-294 of SEQ ID NO:119, or nucleotides 1-172 of SEQ ID NO:120. ### Claim 18 (depends on 1) The method of claim 1, wherein the nucleic acid sequence of the one or more inhibitory nucleic acid molecules is sufficiently complementary to a sequence within or overlapping with: nucleotides 1-14 of SEQ ID NO:118, or nucleotides 1-294 of SEQ ID NO:119, or nucleotides 1-172 of SEQ ID NO:120 to direct target-specific RNA interference (RNAi).