INHIBITION OF GENE EXPRESSION BY NORMAL & MODIFIED ANTISENSE OLIGONUCLEOTIDES
INHIBITION OF GENE EXPRESSION BY NORMAL & MODIFIED ANTISENSE OLIGONUCLEOTIDES
批准号:
3792518
负责人:
C J MARCUS-SEKURA
金额:
$0.0万
依托单位:
--
依托单位国家:
美国
项目类别:
财政年份:
--
资助国家:
美国
项目状态:
未结题
起止时间:
至
关键词:
Epstein Barr virus antisense nucleic acid antiviral agents chloramphenicol acetyltransferase enzyme activity enzyme linked immunosorbent assay gene expression genetic regulation high performance liquid chromatography immunofluorescence technique monoclonal antibody nucleic acid chemical synthesis nucleic acid sequence oligonucleotides phorbols protein purification tissue /cell culture transfection virus antigen
中文摘要
该项目最初是为了确定是否
适当反义序列的修饰寡核苷酸可以抑制
生物反应,并检查其作为抗病毒药物的潜力
剂. 真核细胞中的瞬时表达系统用于
看看正常和修饰的寡核苷酸是否能抑制
氯霉素转移酶CAT(CAT的基因)的活性
通过DNA转染引入细胞)。 认定
微摩尔浓度的修饰寡核苷酸可以
抑制CAT活性和链长、浓度和性质
寡核苷酸的化学修饰可以影响
抑制程度。 这部作品已经出版(1987年),
第二篇论文讨论了使用反义寡核苷酸的技术
已出版(1988)。 使用反义寡核苷酸的实验
现在已经扩展到使用EBV研究潜在的抗病毒作用
基因表达作为一个模型。 反义分子似乎抑制
特异性EBV早期抗原EA(D)的表达,
用佛波醇酯TPA刺激B 95 -8细胞,
免疫荧光和ELISA使用EA的单克隆抗体(D)。
另外的反义序列正在检查中,以确定
这种抑制的特异性。 我们购买了一种仪器,
为这些实验合成寡核苷酸,
最近加入了实验室,这将大大有助于供应
这些材料是这个项目的一个限制因素。 我们有
开发了大规模合成反义核酸的方案,
寡核苷酸和HPLC纯化,并计划实施
在近期 目前,这一领域的积极研究已经
由于时间和人员的限制,
英文摘要
This project was initially undertaken in order to determine whether
modified oligonucleotides of appropriate antisense sequence can inhibit
biological reactions, and to examine their potential as antiviral
agents. A transient expression system in eukaryotic cells was used to
see whether normal and modified oligonucleotides could inhibit the
activity of the enzyme chloramphenicol transferase CAT (the gene for CAT
was introduced into the cells by DNA transfection). It was determined
that micromolar concentrations of modified oligonucleotides could
inhibit CAT activity and that chain length, concentration, and nature of
the chemical modification of the oligonucleotide could influence the
degree of inhibition. This work has been published (1987) as well as a
second paper discussing techniques for using antisense oligonucleotides
has been published (1988). Experiments using antisense oligonucleotides
have now been extended to study potential antiviral effects using EBV
gene expression as a model. Antisense molecules appear to inhibit
expression of a specific EBV early antigen, EA(D), induced by
stimulation of B95-8 cells with the phorbol ester TPA as assayed by
immunofluorescence and ELISA using a monoclonal antibody to EA(D).
Additional antisense sequences are being examined to determine the
specificity of this inhibition. We have purchased an instrument to
synthesize oligonucleotides for these experiments and J. Smith has
recently joined the laboratory, which should help greatly to supply
materials which have been a limiting factor in this project. We have
developed protocols for large-scale synthesis of antisense
oligonucleotides and purification by HPLC, and implementation is planned
in the near future. Temporarily, active research in this area has been
curtailed due to time and personnel constraints.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
IMMUNOLOGIC ANALYSIS OF A HERPES VIRUS GLYCOPROTEIN VACCINE CANDIDATE--EBV GP350
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批准号:3770329
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项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:C J MARCUS-SEKURA
-
依托单位:--
EXPRESSION OF HIV ANTIGENS IN E COLI
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批准号:3811240
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项目类别:
-
资助金额:$0.0万
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财政年份:--
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负责人:C J MARCUS-SEKURA
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依托单位:--
DETECTION OF VIRUSES IN CELL SUBSTRATES
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批准号:3792523
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:C J MARCUS-SEKURA
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依托单位:--
DETECTION OF VIRUSES IN CELL SUBSTRATES
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批准号:3804796
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:C J MARCUS-SEKURA
-
依托单位:--
EXPRESSION OF HIV ANTIGENS IN E COLI
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批准号:3804789
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项目类别:
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资助金额:$0.0万
-
财政年份:--
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负责人:C J MARCUS-SEKURA
-
依托单位:--
INHIBITION OF GENE EXPRESSION BY NORMAL & MODIFIED ANTISENSE OLIGONUCLEOTIDES
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批准号:3804790
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项目类别:
-
资助金额:$0.0万
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财政年份:--
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负责人:C J MARCUS-SEKURA
-
依托单位:--
EPITOPE MAPPING OF THE EBV MEMBRANE ANTIGEN
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批准号:3804788
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:C J MARCUS-SEKURA
-
依托单位:--
EXPRESSION OF HIV ANTIGENS IN E COLI
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批准号:3792517
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项目类别:
-
资助金额:$0.0万
-
财政年份:--
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负责人:C J MARCUS-SEKURA
-
依托单位:--
ANALYSIS OF THE HIV INTEGRATION REACTION USING RECOMBINANT INTEGRASE
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批准号:3770330
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:C J MARCUS-SEKURA
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依托单位:--
DETECTION OF VIRUSES IN CELL LINES USED TO MANUFACTURE BIOLOGICAL PRODUCTS
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批准号:3770331
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:C J MARCUS-SEKURA
-
依托单位:--
ANALYSIS OF THE HIV INTEGRATION REACTION USING RECOMBINANT INTEGRASE
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批准号:3748159
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项目类别:
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资助金额:$0.0万
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财政年份:--
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负责人:C J MARCUS-SEKURA
-
依托单位:--
EPITOPE MAPPING OF THE EBV MEMBRANE ANTIGEN
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批准号:3811239
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项目类别:
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资助金额:$0.0万
-
财政年份:--
-
负责人:C J MARCUS-SEKURA
-
依托单位:--
INHIBITION OF GENE EXPRESSION BY NORMAL & MODIFIED ANTISENSE OLIGONUCLEOTIDES
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批准号:3811241
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项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:C J MARCUS-SEKURA
-
依托单位:--
EPITOPE MAPPING OF THE EBV MEMBRANE ANTIGEN
-
批准号:3792516
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项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:C J MARCUS-SEKURA
-
依托单位:--
DETECTION OF VIRUSES IN CELL SUBSTRATES
-
批准号:3811247
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项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:C J MARCUS-SEKURA
-
依托单位:--
海外基金