Cannabinoid 2 (CB
Cannabinoid receptor 2 (CB 2 ) is expressed in B lymphocytes and is involved in immune regulation. Mouse splenic B cells express three CB 2 transcripts utilizing two different first exons. Human peripheral blood B cells express one CB 2 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 CB 2 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 CB 2 gene (e.g., RNAi molecules).
1. A method of reducing the expression of a B cell CB 2 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 CB 2 receptor gene promoter region within the cell, thereby reducing expression of the B cell CB 2 receptor gene.
2. 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 CB 2 receptor gene.
3. 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.
4. 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.
5. 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.
6. The method of claim 5 , wherein said administering results in regulation of an immunoglobulin E (IgE)-mediated allergic response in the subject.
7. The method of claim 1 , wherein the cell is a human or mouse cell.
8. The method of claim 1 , wherein the one or more inhibitory nucleic acid molecules are contacted with the cell in vitro.
9. The method of claim 1 , wherein the nucleic acid target sequence comprises a region surrounding and including the 5′ end of human exon 1.
10. 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.
11. The method of claim 1 , wherein the nucleic acid target sequence comprises a region surrounding and including the 5′ end of mouse exon 1b.
12. The method of claim 1 , wherein the nucleic acid target sequence comprises a region surrounding and including the 5′ end of exon 2.
13. 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.
14. 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.
15. 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).
16. 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.
17. 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.
18. 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).