Programmed cell death and caspase-12
AssigneeThe General Hospital Corporation
InventorsJunying Yuan, Nobuhiro Morishima
Life of this patent
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Abstract
This invention relates to nucleic acids encoding caspase-12, proteins encoded by casp-12 nucleic acids, vectors, and hosts transformed with vectors. Caspase-12 is a member of the caspase family, which also includes mammalian interleukin-1β-converting enzyme (ICE). Caspases are thought to carry out the execution phase of apoptosis in a cascade of proteolytic events. The invention also relates to the use of caspase-12 to induce programmed cell death.
Claims16
An isolated nucleic acid molecule at least 90% identical to a nucleic acid molecule selected from the group consisting of:
- (a) a nucleic acid molecule consisting of a nucleotide sequence encoding the amino acid sequence of caspase-12S as shown in FIG. 9 (SEQ ID NO:
- 14);
- (b) a nucleic acid molecule consisting of a nucleotide sequence encoding the amino acid sequence of caspase-12S as encoded by the cDNA clone contained in ATCC Deposit No. 209710;
- (c) a nucleic acid molecule consisting of the nucleotide sequence of caspase-12S as shown in FIG. 9 (SEQ ID NO:13);
- (d) a nucleic acid molecule consisting of a nucleotide sequence encoding caspase-12Δ (amino acid residues 95 to 419 of SEQ ID NO:2);
- and
- (e) a nucleic acid molecule consisting of a nucleotide sequence encoding caspase-12Δ2 (amino acid residues 145 to 419 of SEQ ID NO:2),
- wherein said nucleic acid encodes a polypeptide that increases apoptosis.
An isolated nucleic acid molecule encoding a caspase-12S polypeptide, said nucleic acid molecule prepared by a process comprising:
- (a) hybridizing a population of nucleic acid molecules to a nucleic acid molecule comprising the sense or antisense nucleotide sequence of SEQ ID NO:13, wherein the hybridization is performed under stringent hybridization conditions, wherein said stringent hybridization conditions comprise:
- (i) prehybridizing;
- (ii) hybridizing overnight at 65° C. in the hybridization buffer;
- and
- (iii) washing once for 15 minutes in 2×SSC at room temperature, then two times for 30 minutes in 0.1×SSC and 0.1% SDS at 60° C.,
- (b) selecting those nucleic acid molecules of said population that hybridize to said nucleic acid molecule comprising the sense or antisense nucleotide sequence of SEQ ID NO:13;
- and
- (c) selecting nucleic acid molecules of (b) that encode caspase-12S,
- wherein said nucleic acid encodes a polypeptide that increases apoptosis.
A nucleic acid construct comprising a nucleic acid molecule encoding a caspase-12S polypeptide operably linked to a heterologous promoter, wherein said caspase-12S polypeptide is encoded by a nucleic acid molecule at least 90% identical to a nucleic acid molecule selected from the group consisting of:
- (a) a nucleic acid molecule consisting of a nucleotide sequence encoding the amino acid sequence of caspase-12S as shown in FIG. 9 (SEQ ID NO:14);
- (b) a nucleic acid molecule consisting of a nucleotide sequence encoding the amino acid sequence of caspasel2S as encoded by the cDNA clone contained in ATCC Deposit No. 209710;
- and
- (c) a nucleic acid molecule consisting of the nucleotide sequence of caspase-12S as shown in FIG. 9 (SEQ ID NO:13);
- wherein said nucleic acid encodes a polypeptide that increases apoptosis.
A nucleic acid construct comprising a nucleic acid molecule encoding a caspase-12Δ polypeptide operably linked to a heterologous promoter, wherein said caspase-12Δ polypeptide is encoded by a nucleic acid molecule at least 90% identical to a nucleic acid molecule consisting of a nucleotide sequence encoding the amino acid residues 95 to 419 of SEQ ID NO:14 and encodes a polypeptide that increases apoptosis.
A nucleic acid construct comprising a nucleic acid molecule encoding a caspase-12Δ2 polypeptide operably linked to a heterologous promoter, wherein said caspase-12Δ2 polypeptide is encoded by a nucleic acid molecule at least 90% identical to a nucleic acid molecule consisting of a nucleotide sequence encoding the amino acid residues 145 to 419 of SEQ ID NO:14 and encodes a polypeptide that increases apoptosis.
Dependencies are inferred from the claim text.