JP2000139464A - ANTISENSE NUCLEIC ACID COMPOUND FOR INHIBITING EXPRESSION OF p300 OR CBP - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an antisense nucleic acid compound inhibiting the genes encoding p300 and CBP and useful for medicines for treating and diagnosing leukemia, or the like, reagents for analyzing the functions of genes, or the like, by disposing a specific base sequence. SOLUTION: This antisense nucleic acid compound for inhibiting the expression of p300 or CBP has base sequences of formulas I to V or their substantially same base sequences. The antisence nucleic acid compound for inhibiting the expression of the p300 or CBP preferably comprises a natural type or phosphothioate type oligodeoxyribonucleotide.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an adenovirus E1A binding protein referred to as p300 and a CREBb homologous to p300 and referred to as CBP or CREBBP.
The present invention relates to an antisense nucleic acid compound that inhibits expression of an inding protein, and the antisense nucleic acid compound is effective as a therapeutic agent, a diagnostic agent, and a research reagent. Here, CREB is cAMP response element bin
means ding protein.

[0002]

2. Description of the Related Art p300 and CBP have a function as a coactivator for mediating a basic transcription factor and an enhancer binding factor, and serve as genes for genes involved in signal transduction, development, differentiation, carcinogenesis, apoptosis, immunity and the like. It has been clarified that it is involved in the expression (Hiroaki Kawasaki, Kazuhisa Yokoyama, "Ayumi of Medicine", Vol. 186, No. 10, pp. 694-698 (1
998); RH Giles et al. "TIG", Vol. 14, pp. 178-183 (199)
8); H. Kawasaki et al., Nature, 393, 284-289 (1998)).
As a method for selectively suppressing the expression of these genes,
Methods using ribozymes have been reported (H. Kawasaki
Et al., Nature, 393, 284-289 (1998)). This method is a method of preparing a vector that expresses a ribozyme for a target gene, introducing it into a cell, and expressing the ribozyme for the target gene in a cell, thereby inhibiting the expression of the target gene. is there. However, in this method, it is necessary to design a ribozyme and prepare it in a form in which it is incorporated into a vector, and further, it is necessary to introduce it into a cell and express it. That is, many steps are required to achieve the purpose, and considerable time is required. Furthermore, in order to prepare such a ribozyme-incorporated vector, it is necessary to rely on biological techniques, and it is therefore difficult to easily handle large quantities.

[0003]

Therefore, in order to solve these difficulties, if the production of p300 and CBP can be inhibited by the antisense nucleic acid method instead of the ribozyme method, p300 and CBP can be more simply used. Expression can be controlled. In the antisense nucleic acid method, it is important to find an antisense nucleobase sequence that inhibits the expression of the gene of interest.
The purpose is to find an antisense nucleobase sequence that inhibits the production of 00 and CBP.

[0004]

Means for Solving the Problems The present inventors have conducted intensive studies based on the above-mentioned problems, and as a result, have obtained p300 and CB.
With respect to the gene encoding P, an effective antisense nucleic acid is searched for, and a complementary base sequence that efficiently inhibits the production of the protein, that is, an effective antisense nucleic acid compound is found. And completed the present invention. That is, the present invention relates to an antisense nucleic acid compound for inhibiting expression of p300 or CBP, which has the nucleotide sequence represented by SEQ ID NOS: 1 to 5 or a nucleotide sequence substantially identical thereto.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.
The nucleotide sequence represented by SEQ ID NOS: 1 to 5 or a nucleotide sequence substantially identical thereto is a nucleotide sequence complementary to or substantially complementary to the nucleotide sequence of the gene encoding p300 or CBP. Yes, and p300 or C
1 is a base sequence of an antisense nucleic acid compound having an action of inhibiting BP expression. In addition, having the nucleotide sequences represented by SEQ ID NOS: 1 to 5 or having substantially the same nucleotide sequence means that 1) the nucleotide sequence itself represented by SEQ ID NOS: 1 to 5; 2) SEQ ID NOS: 1 to 5 Wherein T is converted to U in the nucleotide sequence represented by: 3) having at least 70% homology with 1) or 2), preferably having at least 80% homology, more preferably Those having a homology of 90% or more, 4) those having a base sequence of p300 or CBP to which a base is added to the base sequence of 1) to 3), and 5) 1) to not more than about 30 bases; 3) a part of the base sequence is deleted from the base sequence, and the base number is 10 or more, preferably 14 or more;
More preferably 17 or more, p300 or CB
It means those having an effect of inhibiting the expression of P.
Here, “gene encoding p300 or CBP”
Are structural genes that define the amino acid sequence of p300 or CBP, and intervening sequences that are present in the middle of the structural genes.
(Intron) and the nucleotide sequence upstream of a structural gene (such as a promoter or operator) or the nucleotide sequence downstream of a structural gene (such as polyA) involved in the expression of the factor. Also,
The antisense nucleic acid compound of the present invention has p300 or C
BP expression is significantly inhibited as compared with the control, and BP expression is inhibited by 70% or less, 50% or less by effective nucleotides,
% Or less particularly effectively. The inhibitory effects of these antisense nucleic acid compounds can be confirmed in cultured cell lines. Here, “inhibiting the expression of p300 or CBP” means “mRNA encoding p300 or CBP”.
Inhibits the translation of p300 into CBP or CBP, or the like, means that the protein is not produced or suppressed.

[0006] The antisense nucleic acid compound of the present invention is designed based on the concept of the antisense nucleic acid method as follows, prepared based on the antisense nucleic acid method, and produced the protein p300 or CBP which produces its effect. It was obtained by evaluation based on the amount. That is, first, p300 or CB
Obtain nucleotide sequence information of the gene encoding P. Next, among nucleic acid compounds complementary to the partial base sequence of the gene, a nucleic acid compound expected to have an effect as an antisense nucleic acid is selected, and the selected antisense nucleic acid is prepared by a synthetic chemical method or the like. did. Whether the compound thus obtained efficiently inhibits the production of p300 or CBP was tested using a cultured cell line, and p3
A nucleic acid compound containing a base sequence that inhibits the production of 00 or CBP is the antisense nucleic acid compound of the present invention. Note that, in the present invention, expression is considered to be inhibited when the inhibition rate of p300 or CBP expression by the synthesized nucleic acid compound is greater than the p300 or CBP expression inhibition rate by a nucleic acid compound having a “random base sequence”. It is. The “random base sequence” refers to a base sequence in which the degree of complementarity to the gene encoding the target protein is statistically expected to be lower than or equal to the degree of complementarity. The antisense nucleic acid compound of the present invention can significantly suppress the expression of p300 or CBP, but preferred for the present invention are those that inhibit the expression of p300 or CBP to 70% or less, and more preferable ones. Is 50% or less, and particularly preferred is 30% or less.

O Antisense nucleic acid method (1) Selection of antisense nucleic acid Obtain the nucleotide sequences of proteins such as p300 and CBP. This is, as information described in the paper, GenB
The information can be obtained as information stored in a gene database such as ank, or as a result of directly analyzing a nucleotide sequence. Next, among nucleic acid compounds complementary to the partial base sequence of the gene, a nucleic acid compound expected to have an effect as an antisense nucleic acid is selected. As this method, mRNA encoding the target protein is used.
Alternatively, a method based on the prediction of the secondary structure of the precursor (Kiyoshi Uchida, Naoki Sugimoto, Antisense 1: 6-11 (1997)), searching for the possibility of forming a duplex, A method to select from difficult areas (JP-A-10-52285; Kiyoshi Uchida, Naoki Sugimoto, A
ntisense 1 6-11 (1997); Uchida Kiyoshi, Chemistry and Industry 50
1758-1761 (1967); Kiyoshi Uchida, Annual Report of Toagosei Research TR
END First issue 52-56 (1998); Uchida Kiyoshi Antisense Volume 2
3-7 (1998)), and a method of selecting from near the start of translation (Kiyoshi Uchida, Naoki Sugimoto, Antisense 1, 6-11 (1997)). (2) Preparation of nucleic acid compound The nucleic acid compound includes natural oligodeoxyribonucleotides, phosphorothioate-type oligodeoxyribonucleotides, phosphorodithioate-type oligodeoxyribonucleotides, methylphosphonate-type oligodeoxyribonucleotides, and phosphoramidate-type oligodeoxyribonucleotides. Oligodeoxyribonucleotides, H-phosphonate-type oligodeoxyribonucleotides, triester-type oligodeoxyribonucleotides, α-anomeric oligodeoxyribonucleotides, oligoribonucleotides corresponding to each of the above-mentioned oligodeoxyribonucleotides, peptide nucleic acids, and other artificial Nucleic acids and nucleic acid-modifying compounds can be mentioned. Natural-type and phosphorothioate-type oligodeoxyribonucleotides are represented by mR NA
Is a preferred nucleic acid compound in that it becomes a substrate for RNase H when it forms a double chain by binding to, and has little nonspecific expression inhibition and easy synthesis. Synthesis of a nucleic acid compound of a natural type can be performed, for example, by using ABI (Applied Biosystems In
c.) Using the 381A DNA synthesizer manufactured by the company or the 394 DNA / RNA synthesizer manufactured by the company, the phosphoramidite method (ABI's procedure manual, or Oligonucleotides and Analyte, edited by F. Eckstein)
ogues: A Practical Approach, see IRL Press, 1991). The phosphoramidite method is a method in which a modified deoxyribonucleoside or a modified ribonucleoside is bound to a phosphoramidite protected with a cyanoethyl group or the like at the 3 ′ end of the modified deoxyribonucleoside, another modified deoxyribonucleoside, a modified ribonucleoside, an oligo-modified deoxyribonucleotide. This is a method for synthesizing nucleic acid-related compounds such as oligodeoxyribonucleotides and oligoribonucleotides, which is based on condensation with the 5 ′ end of oligo-modified ribonucleotides. Then, in the final cycle of the synthesis, the protecting group for the sugar hydroxyl group at the 5′-terminal is
(Synthesis is completed in a state where dimethoxytrityl group or the like is bonded). After the oligomer synthesized at room temperature is cleaved from the support, the base portion and the phosphate portion are deprotected. Thus, a crude product of a natural oligonucleic acid is obtained. Phosphorothioate-type nucleic acid compounds are AB
It can be synthesized by the phosphoramidite method using a synthesizer of Company I. The processing after the final cycle of the synthesis is the same as in the case of the natural type described above. The resulting crude product of the nucleic acid compound can be purified by a conventional purification method, for example, ethanol precipitation, or reversed-phase chromatography, ion-exchange chromatography, and high-performance liquid chromatography based on gel filtration chromatography. (HPL
C), various chromatography such as supercritical chromatography, and further, electrophoresis can be used for purification. In addition, cartridges manufactured based on the principle of reversed-phase chromatography [for example, Seppak Plus (filled with tC18) (Longbody / EN
V); Waters]. Phosphorothioate-type nucleic acid compound (20-mer, about 3 mg in crude product)
Is the same as in the case of the above-mentioned natural type. The purity of the natural type and phosphorothioate type nucleic acid compounds is as follows:
It can be examined by HPLC analysis or capillary electric swimming course. The synthesized nucleic acid compound is used in an evaluation experiment using a cultured cell system described below. (3) Evaluation Experiment in Cultured Cell System An evaluation experiment of a nucleic acid compound can be performed using a culture cell system. That is, the effect on the expression of p300 or CBP is examined in a cultured cell system using an antisense nucleic acid compound such as a phosphorothioate type oligodeoxyribonucleotide or a natural type oligodeoxyribonucleotide having a nucleotide sequence expected to have an antisense nucleic acid effect. be able to. That is, under sterile conditions, human, mouse, rat, guinea pigs, cells derived from mammals such as cattle, antisense nucleic acid compounds such as phosphorothioate-type oligodeoxyribonucleotides or natural-type oligodeoxyribonucleotides
0.01-100 μM, preferably 0.1-10 μM, if necessary, lipofectin reagent, lipofectamine reagent, DO
TAP reagent, Tfx reagent, artificial synthetic lipid vesicle, liposome, membrane fusion reagent, polymeric micelle-forming reagent, polymer carrier, or other intracellular transfection reagents, thereby inhibiting the expression of the target protein by p300 or CBP ELISA or Western blotting using an antibody against mR, or mR encoding p300 or CBP
By examining NA by RT-PCR (reverse transcriptase-polymerase chain reaction) or the like, the protein expression inhibitory effect of the used antisense nucleic acid compound can be confirmed.

[0008]

[Example 1]

(1) Selection of antisense nucleic acid First, the nucleotide sequences of p300 and CBP mRNA were obtained from GenBank, a public gene database. Next, the possibility of forming a double strand was searched, and an antisense nucleic acid having a complementary base sequence was selected for a region in which a double strand was hardly formed by using the previously proposed method (Japanese Patent Laid-Open No. −52285; Uchida Kiyoshi, Sugimoto Naoki, Antisense
1, 6-11 (1997); Uchida Kiyoshi, Chemistry and Industry
50, 1758-1761 (1997); Toagosei Kenkyu Kenkyu TREND, first issue, 52-56 (199)
8); Uchida, Antisense 2, 3-7 (199)
8)). That is, for p300, as a result of searching for the base sequence represented by SEQ ID NO: 6, the following five nucleic acid compounds were selected as antisense nucleic acid compound candidates. <Nucleic acid compound name><Basesequence><SEQ ID NO> A0286U CATCTGGTAAGTCATGTTCCA (SEQ ID NO: 1) A0662T CATCCCTGTGTTCATTCCCA (SEQ ID NO: 2) A1357X TTTTCCAGTGTGAAATGATTTGTC (SEQ ID NO: 3) A1586W GAATCTGGTTTACTTGATAGGGT (SEQ ID NO: 7) As a result of a search for the base sequence represented by SEQ ID NO: 9 for CBP, the following five nucleic acid compounds were selected as antisense nucleic acid compound candidates. <Nucleic acid compound name><Basesequence><SEQ ID NO> A0450U TGCTTTTGTGCTTGTGGATTC (SEQ ID NO: 4) A1978X CTTTTCTTCTAGTTCTTTTTGTATAT (SEQ ID NO: 10) A3159U GCTTCCACTTTTACTTCAGGT (SEQ ID NO: 11) A4674U TTCCTTTCTTCTTCTTCTTGT (SEQ ID NO: 5TGTTCTGTTTCTGTCT2GTTTCTGTGTGTCT2GTTTGTGTGTCTCTGTGTGTCT1GTTG1TGTTCTGTGTCTCTGTGTGTGTCTCTGTGTCTTGTTCTCTGTGTGTCTCTGTGTCTTGTTCTGTGTCTTGTTCTGTGTGTCTCTGTGTGTCTCTTGTTCTGTGT ) Preparation of Nucleic Acid Compound A phosphorothioate-type nucleic acid compound corresponding to the above sequence was prepared by the phosphoramidite method. (3) Evaluation experiment using cultured cell line F9 cells, which are mouse embryonic tumor cells, were prepared using a 6-well plate, and Dulbecco containing 10% fetal calf serum (FCS).
The cells were cultured in co's modified Eagle's medium (DMEM). The number of cells immediately before use in the experiment was 1 × 10 5 cells / well. Antisense nucleic acid is Lipofe
Using ctAMINETM (GibcoBRL) reagent, final concentration 5μ
M was used. That is, a complex of the antisense nucleic acid and LipofectAMINETM was obtained by adding 1 μl of LipofectAMINETM per 1 μg of the antisense nucleic acid and leaving the mixture at room temperature for 20 minutes. The thus obtained complex of antisense nucleic acid and LipofectAMINETM was added to 10
DMEM containing% FCS was used as a medium for monitoring the antisense nucleic acid effect, and was replaced every 24 hours. After 72 hours or 96 hours, proteins were extracted from the cells, and amplified sandwich ELISA (amplified sand ELISA) was performed.
The effect of adding the nucleic acid compound was confirmed by a wich enzyme-linked immunosorbent assay (hereinafter simply referred to as ELISA). The ELISA method used here is ELAST ELISA Amp
The lification system (Du Pont) was used and the protocol provided by the company was followed. That is, an ELISA plate is coated with a monoclonal antibody against p300 (RW128) or a monoclonal antibody against CBP (AC238) (PBS (phosphate-buffered sa).
line) Using buffer solution, 4 ° C, 16 hours), 0.05% Tween
After washing three times with PBS (200 μl) containing 2% B
Blocked with SA (buffer PBS) (2 to 3 hours at room temperature). The samples to be subjected to the ELISA analysis were then systematically diluted, added to the plates prepared as described above, and left at room temperature for 4 hours. This is
Washed three times with PBS containing 05% Tween in the same manner as described above. To this, the above-mentioned antibody specific to p300 or CBP was added and left at room temperature for 1.5 hours. This was washed three times in the same manner as described above, and a biotinylated anti-mouse IgG antibody was added.
Further, streptavidin conjugated with horseradish peroxidase was added (room temperature). After adding phenylenediamine (1.2 mM) as a substrate, the absorbance at 490 nm derived from the reaction product generated from the substrate was followed by a microplate reader. As a result, in p300, an effect was observed in A0286U, A0662T, and A1357X. In particular, A1357X had a remarkable effect (FIG. 1). In FIG. 1, WT is the case where no nucleic acid compound is added, and Random Oligomer is N23 (where N is
A, G, C, T means a mixture of all bases). In CBP, an effect was observed with A0450U and A4674U. Particularly, A0450U had a remarkable effect (FIG. 2). In FIG. 2, WT is the case where no nucleic acid compound was added, and
What is described as Oligomer is N23 (where N means a mixture of all A, G, C, T bases).

[0009]

According to the present invention, there is provided an antisense nucleic acid compound that inhibits the expression of genes encoding p300 and CBP. The antisense nucleic acid compound of the present invention inhibits the production of p300 or CBP.
It is useful for functional analysis of 00 and CBP. Therefore, it can be used for research as an inhibitory reagent for gene function analysis. Also, p300 and CBP are MOZ (Monocytic
Zinc Finger Protein), MLL (Mixed Linear Leukem)
ia), and AML (Acute Myelogeneous Leukemia)
And MOZ-CBP, MLL-CBP, and MLL
The antisense nucleic acid compound of the present invention can also be used as a therapeutic agent for leukemia, for example, because it produces chimeric molecules such as p300 and causes blood-related malignancies. Further, by using the antisense nucleic acid compound of the present invention, differentiation, proliferation, cell cycle, apoptosis can be detected, and a diagnostic agent targeting histone acetyltransferase or transcription coactivator can be developed.

[0010]

[Sequence list] SEQ ID NO: 1 Sequence length: 21 Sequence type: Nucleic acid Number of strands: Single stranded Topology: Linear Sequence type: Antisense nucleic acid Sequence: CATCTGGTAA GTCATGTTCC A SEQ ID NO: 2 Sequence length Length: 20 Sequence type: Nucleic acid Number of strands: Single-stranded Topology: Linear Sequence type: Antisense nucleic acid Sequence: CATCCCTGTG TTCATTCCCA SEQ ID NO: 3 Sequence length: 24 Sequence type: Nucleic acid Number of strands: Single-stranded topology: Linear Sequence type: Antisense nucleic acid Sequence: TTTTCCAGTG TGAAATGATT TGTC SEQ ID NO: 4 Sequence length: 21 Sequence type: Nucleic acid Number of strands: Single-stranded Topology: Linear Sequence type : Antisense nucleic acid Sequence: TGCTTTTGTG CTTGTGGATT C SEQ ID NO: 5 Sequence length: 21 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: Antisense nucleic acid Sequence: TTCCTTTCTT CTTCTTCTTG T SEQ ID NO: 6 Length of sequence: 1743 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: cDNA to mRNA Sequence: GAATTCCGGT CACCGCAACC AGGCGAAGAG AAAAAGGAAC TTCCCCCACC CCTGTTGGTG CGGCGGGGAGCT CCGGAGCC CGGGGCTTGATCGACGTCGACGTTCGTTCGTCGATCTGCTTTCGTCGACGTTCGTT AAATGGCCGA GAATGTGGTG GAACCCGGGC CGCCTTCAGC AAGCGCCTTA AACTCTCATC TCCGGCCCTC TCGGCGTCCG CCAGCGATGG CACAGATTTT GGTTCACTGT TGACCTGGAA CATGACTTAC CAGATGAATT AATCAACTCT ACAGAATTGG GACTAACCAA TGGTGGCGAT AGCAGTCAGC TTCAGACAAG TCTTGGCATA GTACAAGATG CAGCCTCGAA ACATAAACAG CTGTCAGAAC TTGCTGAGGT CTGGTAGCTC CCCAAACCTC AACATGGGAG TCGGTGGCCC AGGCCAAGCG ATGGCCAGCC AGGCCCAACA GAACAGCCCT GGATTAAGTT TGATAAATAG CATGGTCAAA AGCCCAATGG CACAGACAGG CTTGACTTCT CCAAACATGG GGATTGGCAG TAGTGGACCA AATCAGGGTC CTACTCAGTC CCCAGCAGGT ATGATGAACA GTCCAGTGAA CCAGCCTGCC ATGGGAATGA ACACAGGGAT GAATGCTGGC ATGAATCCTG GAATGTTGGC TGCAGGCAAT GGACAAGGGA TAATGCCCAA TCAAGTCATG AACGGTTCCA TTGGAGCAGG CC GGGGACGG CCAAACATGC AGTACCCAAA TGCAGGCATG GGCAATGCTG GCAGTTTATT GACTGAGCCA CTACAGCAGG GCTCTCCTCA GATGGGAGGA CAGCCAGGAT TGAGAGGCCC CCAACCACTT AAGATGGGAA TGATGAACAA TCCCAGTCCT TATGGTTCAC CATACACTCA GAATTCTGGA CAGCAGATTG GAGCAAGTGC CTTGGTCTCC AAATTCAGAC AAAGACTGTT CTACCAAATA ACTTATCTCC ATTTGCAATG GACAAAAAGC AGTTCCTGGT GGGGGAATGC CCAGTATGGG CCAGCAGCCT ACCCATCGGT CCAGCAGCCA GGCCTGGACT CCAGTTGCCG CAGGAATGGG TTCTGGAGCA CACACAGCTG ATCCAGAGAA GCGCAACGTC ATCCCAGCAG CTTGTTCTCC TTTTACATGC TCACAAGTGC CAGCGCCGGG AGCAAGCTAA TGGGGAAGTG AGGCAGTGCA ACCTTCCTCA CTGTCGTACC ATGAAAAATG TCCTAAACCA TATGACACAT TGCCAGTCAG GCAAATCCTG CCAAGTGGCA CATTGTGCAT CTTCTCGACA AATCATTTCA CACTGGAAAA ATTGCACAAG GCATGATTGT CCTGTGTGTC TTCCTCTCAA AAATGCTGGG GATAAGCGAA ATCAACAGTC AATTTTGACT GGAGCACCAG TTGGGCTTGG AAACCCTAGC TCTCTAGGAG TGGGGCAGCA GTCCACTCCT AGCCTAAGCA CTGTTAGTCA GATTGACCCC AGCTCTATAG AGCGAGCTTA CGCTGCTCTT GGACTACCCT ATCAAGTAAA CCAGATTCCA CCACAACCCC AGGTACAAGC AAAGAATCAA CAAAGCCAGC CATCTGGACA GTCTCCCCAG GGGCATGCGG T CTGTGAACA ACATGAGTGC TAGTCCTATG GGTGTAAATG GAGGTGTTGG GGTTCAGACG CCA SEQ ID NO: 7 Sequence length: 23 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: Antisense nucleic acid Sequence: GAATCTGGTT TACTTGATAG GGT SEQ ID NO: 8 Sequence length: 22 Sequence type: Nucleic acid Number of strands: Single stranded Topology: Linear Sequence type: Antisense nucleic acid Sequence: TTGTTGATTC TTTGCTTGTA CC SEQ ID NO: 9 Sequence length: 7326 Sequence type: Nucleic acid number of strands: single strand topology: linear sequence type: cDNA-to mRNA sequences: ATGGCCGAGA ACTTGCTGGA CGGACCGCCC AACCCCAAAC GAGCCAAACT CAGCTCGCCC GGCTTCTCCG CGAATGACAA CACAGATTTT GGATCATTGT TTGACTTGGA AAATGACCTT CCTGATGAGC TGATCCCCAA TGGAGAATTA AGCCTTTTAA ACAGTGGGAA CCTTGTTCCA GATGCTGCGT CCAAACATAA ACAACTGTCA GAGCTTCTTA GAGGAGGCAG CGGCTCTAGC ATCAACCCAG GGATAGGCAA TGTGAGTGCC AGCAGCCCTG TGCAACAGGG CCTTGGTGGC CAGGCTCAGG GGCAGCCGAA CAGT ACAAAC ATGGCCAGCT TAGGTGCCAT GGGCAAGAGC CCTCTGAACC AAGGAGACTC ATCAACACCC AACCTGCCCA AACAGGCAGC CAGCACCTCT GGGCCCACTC CCCCTGCCTC CCAAGCACTG AATCCACAAG CACAAAAGCA AGTAGGGCTG GTGACCAGTA GTCCTGCCAC ATCACAGACT GGACCTGGGA TCTGCATGAA TGCTAACTTC AACCAGACCC ACCCAGGCCT TCTCAATAGT AACTCTGGCC ATAGCTTAAT GAATCAGGCT CAACAAGGGC AAGCTCAAGT CATGAATGGA TCTCTTGGGG CTGCTGGAAG AGGAAGGGGA GCTGGAATGC CCTACCCTGC TCCAGCCATG CAGGGGGCCA CAAGCAGTGT GCTGGCGGAG ACCTTGACAC AGGTTTCCCC ACAAATGGCT GGCCATGCTG GACTAAATAC AGCACAGGCA GGAGGCATGA CCAAGATGGG AATGACTGGT ACCACAAGTC CATTTGGACA ACCCTTTAGT CAAACTGGAG GGCAGCAGAT GGGAGCCACT GGAGTGAACC CCCAGTTAGC CAGCAAACAG AGCATGGTCA ATAGTTTACC TGCTTTTCCT ACAGATATCA AGAATACTTC AGTCACCACT GTGCCAAATA TGTCCCAGTT GCAAACATCA GTGGGAATTG TACCCACACA AGCAATTGCA ACAGGCCCCA CAGCAGACCC TGAAAAACGC AAACTGATAC AGCAGCAGCT GGTTCTACTG CTTCATGCCC ACAAATGTCA GAGACGAGAG CAAGCAAATG GAGAGGTTCG NGCCTGTTCT CTCCCACACT GTCGAACCAT GAAAAACGTT TTGAATCACA TGACACATTG TCAGGCTCCC AAAGCCTGCC AAGTTGCCCA TTGTGCATCT TCA CGACAAA TCATCTCTCA TTGGAAGAAC TGCACACGAC ATGACTGTCC TGTTTGCCTC CCTTTGAAAA ATGCCAGTGA CAAGCGAAAC CAACAAACCA TCCTGGGATC TCCAGCTAGT GGAATTCAAA ACACAATTGG TTCTGTTGGT GCAGGGCAAC AGAATGCCAC TTCCTTAAGT AACCCAAATC CCATAGACCC CAGTTCCATG CAGCGGGCCT ATGCTGCTCT AGGACTCCCC TACATGAACC AGCCTCAGAC GCAGCTGCAG CCTCAGGTTC CTGGCCAGCA ACCAGCACAG CCTCCAGCCC ACCAGCAGAT GAGGACTCTC AATGCCCTAG GAAACAACCC CATGAGTGTC CCAGCAGGAG GAATAACAAC AGATCAACAG CCACCAAACT TGATTTCAGA ATCAGCTCTT CCAACTTCCT TGGGGGCTAC CAATCCACTG ATGAATGATG GTTCAAACTC TGGTAACATT GGAAGCCTCA GCACGATACC TACAGCAGCG CCTCCTTCCA GCACTGGTGT TCGAAAAGGC TGGCATGAAC ATGTGACTCA GGACCTACGG AGTCATCTAG TCCATAAACT CGTTCAAGCC ATCTTCCCAA CTCCAGACCC TGCAGCTCTG AAAGATCGCC GCATGGAGAA CCTGGTTGCC TATGCTAAGA AAGTGGAGGG AGACATGTAT GAGTCTGCTA ATAGCAGGGA TGAATACTAT CATTTATTAG CAGAGAAAAT CTATAAAATA CAAAAAGAAC TAGAAGAAAA GCGGAGGACA CGTTTACATA AGCAAGGCAT CCTGGGTAAC CAGCCAGCTT TACCAGCTTC TGGGGCTCAG CCCCCTGTGA TTCCACCAGC CCAGTCTGTA AGACCTCCAA ATGGGCCCCT GCCTTTGCCA GTGAATCGCA TG CAGGTTTC TCAAGGGATG AATTCATTTA ACCCAATGTC CCTGGGAAAC GTCCAGTTGC CACAGGCACC CATGGGACCT CGTGCAGCCT CCCCTATGAA CCACTCTGTG CAGATGAACA GCATGGCCTC AGTTCCGGGT ATGGCCATTT CTCCTTCACG GATGCCTCAG CCTCCAAATA TGATGGGCAC TCATGCCAAC AACATTATGG CCCAGGCACC TACTCAGAAC CAGTTTCTGC CACAGAACCA GTTTCCATCA TCCAGTGGGG CAATGAGTGT GAACAGTGTG GGCATGGGGC AACCAGCAGC CCAGGCAGGT GTTTCACAGG GTCAGGAACC TGGAGCTGCT CTCCCTAACC CTCTGAACAT GCTGGCACCC CAGGCCAGCC AGCTGCCTTG CCCACCAGTG ACACAGTCAC CATTGCACCC GACTCCACCT CCTGCTTCCA CAGCTGCTGG CATGCCCTCT CTCCAACATC CAACGGCACC AGGAATGACC CCTCCTCAGC CAGCAGCTCC CACTCAGCCA TCTACTCCTG TGTCATCTGG GCAGACTCCT ACCCCAACTC CTGGCTCAGT GCCCAGCGCT GCCCAAACAC AGAGTACCCC TACAGTCCAG GCAGCAGCAC AGGCTCAGGT GACTCCACAG CCTCAGACCC CAGTGCAGCC ACCATCTGTG GCTACTCCTC AGTCATCACA GCAGCAACCA ACGCCTGTGC ATACTCAGCC ACCTGGCACA CCGCTTTCTC AGGCAGCAGC CAGCATTGAT AATAGAGTCC CTACTCCCTC CACTGTGACC AGTGCTGAAA CCAGTTCCCA GCAGCCAGGA CCCGATGTGC CCATGCTGGA AATGAAGACA GAGGTGCAGA CAGATGATGC TGAGCCTGAA CCTACTGAAT CCAAGGGGGA A CCTCGGTCT GAGATGATGG AAGAGGATTT ACAAGGTTCT TCCCAAGTAA AAGAAGAGAC AGATACGACA GAGCAGAAGT CAGAGCCAAT GGAAGTAGAA GAAAAGAAAC CTGAAGTAAA AGTGGAAGCT AAAGAGGAAG AAGAGAACAG TTCGAACGAC ACAGCCTCAC AATCAACATC TCCTTCCCAG CCACGCAAAA AAATCTTTAA ACCCGAGGAG CTACGCCAGG CACTTATGCC AACTCTAGAA GCACTCTATC GACAGGACCC AGAGTCTTTG CCTTTTCGTC AGCCTGTAGA TCCTCAGCTC CTAGGAATCC CAGATTATTT TGATATAGTG AAGAATCCTA TGGACCTTTC TACCATCAAA CGAAAGCTGG ACACAGGGCA ATATCAAGAA CCCTGGCAGT ATGTGGATGA TGTCAGGCTT ATGTTCAACA ATGCGTGGCT ATATAATCGT AAAACGTCCC GTGTATATAA ATTTTGCAGT AAACTTGCAG AGGTCTTTGA ACAAGAAATT GACCCTGTCA TGCAGTCTCT TGGATATTGC TGTGGACGAA AGTATGAGTT CTCCCCACAG ACTTTGTGCT GTTACGGAAA GCAGCTGTGT ACAATTCCTC GTGATGCAGC CTACTACAGC TATCAGAATA GGTATCATTT CTGTGGGAAG TGTTTCACAG AGATCCAGGG CGAGAATGTG ACCCTGGGTG ACGACCCTTC CCAACCTCAG ACGACAATTT CCAAGGATCA ATTTGAAAAG AAGAAAAATG ATACCTTAGA TCCTGAACCT TTTGTTGACT GCAAAGAGTG TGGCCGGAAG ATGCATCAGA TTTGTGTTCT ACACTATGAC ATCATTTGGC CTTCAGGTTT TGTGTGTGAC AACTGTTTGA AGAAAACTGG CAGACCTCGG AAAGAAAACA AATTCAGTGC TAAGAGGCTG CAGACCACAC GATTGGGAAA CCACTTAGAA GACAGAGTGA ATAAGTTTTT GCGGCGCCAG AATCACCCTG AAGCTGGGGA GGTTTTTGTC AGAGTGGTGG CCAGCTCAGA CAAGACTGTG GAGGTCAAGC CGGGAATGAA GTCAAGGTTT GTGGATTCTG GAGAGATGTC GGAATCTTTC CCATATCGTA CCAAAGCACT CTTTGCTTTT GAGGAGATCG ATGGAGTCGA TGTGTGCTTT TTTGGGATGC ATGTGCAAGA TACGGCTCTG ATTGCCCCCC ACCAAATACA AGGCTGTGTA TACATATCTT ATCTGGACAG TATTCATTTC TTCCGGCCCC GCTGCCTCCG GACAGCTGTT TACCATGAGA TCCTCATCGG ATATCTCGAG TATGTGAAGA AATTGGTGTA TGTGACAGCA CATATTTGGG CCTGTCCCCC AAGTGAAGGA GATGACTATA TCTTTCATTG CCACCCCCCT GACCAGAAAA TCCCCAAACC AAAACGACTA CAGGAGTGGT ACAAGAAGAT GCTGGACAAG GCGTTTGCAG AGAGGATCAT TAACGACTAT AAGGACATCT TCAAACAAGC GAACGAAGAC AGGCTCACGA GTGCCAAGGA GTTGCCCTAT TTTGAAGGAG ATTTCTGGCC TAATGTGTTG GAAGAAAGCA TTAAGGAACT AGAACAAGAA GAAGAAGAAA GGAAAAAAGA AGAGAGTACT GCAGCGAGTG AGACTCCTGA GGGCAGTCAG GGTGACAGCA AAAATGCGAA GAAAAAGAAC AACAAGAAGA CCAACAAAAA CAAAAGCAGC ATTAGCCGCG CCAACAAGAA GAAGCCCAGC ATGCCCAATG TTTCCAACGA CCTGTCGCAG AAGCTGTATG CCACCATGGA GAAGCACAAG GAGGTATTCT TTGTGATTCA TCTGCATGCT GGGCCTGTTA TCAGCACTCA GCCCCCCATC GTGGACCCTG ATCCTCTGCT TAGCTGTGAC CTCATGGATG GGCGAGATGC CTTCCTCACC CTGGCCAGAG ACAAGCACTG GGAATTCTCT TCCTTACGCC GCTCCAAATG GTCCACTCTG TGCATGCTGG TGGAGCTGCA CACACAGGGC CAGGACCGCT TTGTTTATAC CTGCAATGAG TGCAAACACC ATGTGGAAAC ACGCTGGCAC TGCACTGTGT GTGAGGACTA TGACCTTTGT ATCAATTGCT ACAACACAAA GAGCCACACC CATAAGATGG TGAAGTGGGG GCTAGGCCTA GATGATGAGG GCAGCAGTCA GGGTGAGCCA CAGTCCAAGA GCCCCCAGGA ATCCCGGCGT CTCAGCATCC AGCGCTGCAT CCAGTCCCTG GTGCATGCCT GCCAGTGTCG CAATGCCAAC TGCTCACTGC CGTCTTGCCA GAAGATGAAG CGAGTCGTGC AGCACACCAA GGGCTGCAAG CGCAAGACTA ATGGAGGATG CCCAGTGTGC AAGCAGCTCA TTGCTCTTTG CTGCTACCAC GCCAAACACT GCCAAGAAAA TAAATGCCCT GTGCCCTTCT GCCTCAACAT CAAACATAAC GTCCGCCAGC AGCAGATCCA GCACTGCCTG CAGCAGGCTC AGCTCATGCG CCGGCGAATG GCAACCATGA ACACCCGCAA TGTGCCTCAG CAGAGTTTGC CTTCTCCTAC CTCAGCACCA CCCGGGACTC CTACACAGCA GCCCAGCACA CCCCAAACAC CACAGCCCCC AGCCCAGCCT CAGCCTTCAC CTGTTAACAT GTCACCAGCA GGCTTCCCTA ATGTAGCCC G GACTCAGCCC CCAACAATAG TGTCTGCTGG GAAGCCTACC AACCAGGTGC CAGCTCCCCC ACCCCCTGCC CAGCCCCCAC CTGCAGCAGT AGAAGCAGCC CGGCAAATTG AACGTGAGGC CCAGCAGCAG CAGCACCTAT ACCGAGCAAA CATCAACAAT GGCATGCCCC CAGGACGTGA CGGTATGGGG ACCCCAGGAA GCCAAATGAC TCCTGTGGGC CTGAATGTGC CCCGTCCCAA CCAAGTCAGT GGGCCTGTCA TGTCTAGTAT GCCACCTGGG CAGTGGCAGC AGGCACCCAT CCCTCAGCAG CAGCCGATGC CAGGCATGCC CAGGCCTGTA ATGTCCATGC AGGCCCAGGC AGCAGTGGCT GGGCCACGGA TGCCCAATGT GCAGCCAAAC AGGAGCATCT CGCCAAGTGC CCTGCAAGAC CTGCTACGGA CCCTAAAGTC ACCCAGCTCT CCTCAGCAGC AGCAGCAGGT GCTGAACATC CTTAAATCAA ACCCACAGCT AATGGCAGCT TTCATCAAAC AGCGCACAGC CAAGTATGTG GCCAATCAGC CTGGCATGCA GCCCCAGCCC GGACTTCAAT CCCAGCCTGG TATGCAGCCC CAGCCTGGCA TGCACCAGCA GCCTAGTTTG CAAAACCTGA ACGCAATGCA AGCTGGTGTG CCACGGCCTG GTGTGCCTCC ACCACAACCA GCAATGGGAG GCCTGAATCC CCAGGGACAA GCTCTGAACA TCATGAACCC AGGACACAAC CCCAACATGA CAAACATGAA TCCACAGTAC CGAGAAATGG TGAGGAGACA GCTGCTACAG CACCAGCAGC AGCAGCAGCA ACAGCAGCAG CAGCAGCAGC AACAACAAAA TAGTGCCAGC TTGGCCGGGG GCATGGCGGG ACACAGCC AG TTCCAGCAGC CACAAGGACC TGGAGGTTAT GCCCCAGCCA TGCAGCAGCA ACGCATGCAA CAGCACCTCC CCATCCAGGG CAGCTCCATG GGCCAGATGG CTGCTCCAAT GGGACAACTT GGCCAGATGG GGCAGCCTGG GCTAGGGGCA GACAGCACCC CTAATATCCA GCAGGCCCTG CAGCAACGGA TTCTGCAGCA GCAGCAGATG AAGCAACAAA TTGGGTCACC AGGCCAGCCG AACCCCATGA GCCCCCAGCA GCACATGCTC TCAGGACAGC CACAGGCCTC ACATCTCCCT GGCCAGCAGA TCGCCACATC CCTTAGTAAC CAGGTGCGAT CTCCAGCCCC TGTGCAGTCT CCACGGCCCC AATCCCAACC TCCACATTCC AGCCCGTCAC CACGGATACA ACCCCAGCCT TCACCACACC ATGTTTCACC CCAGACTGGA ACCCCTCACC CTGGACTCGC AGTCACCATG GCCAGCTCCA TGGATCAGGG ACACCTGGGG AACCCTGAAC AGAGTGCAAT GCTCCCCCAG CTGAATACCC CCAACAGGAG CGCACTGTCC AGTGAACTGT CCCTGGTTGG TGATACCACG GGAGACACAC TAGAAAAGTT TGTGGAGGGT TTGTAG SEQ ID NO: 10 Sequence length: 24 Sequence type: Nucleic acid strand Sequence type: Nucleic acid type Sequence: Nucleic acid type: : CTTTTCTTCT AGTTCTTTTT GTAT SEQ ID NO: 11 Sequence length: 21 Sequence type: Nucleic acid Number of strands: Single strand Theory: Linear Sequence type: Antisense nucleic acid Sequence: GCTTCCACTT TTACTTCAGG T SEQ ID NO: 12 Sequence length: 21 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: Antisense Nucleic acid sequence: TGTTGGTCTT CTTGTTGTTC T

[Brief description of the drawings]

FIG. 1 shows the effect of nucleic acid compounds on p300 expression.

FIG. 2 shows the effect of a nucleic acid compound on CBP expression.

Claims (1)

[Claims] 1. An antisense nucleic acid compound for inhibiting the expression of p300 or CBP, which has a nucleotide sequence represented by SEQ ID NOS: 1 to 5 or a nucleotide sequence substantially identical thereto.