Rhodopsin Gene Correction and Gene Knockout in Rod Cells
视杆细胞中的视紫红质基因校正和基因敲除
基本信息
- 批准号:7686532
- 负责人:
- 金额:$ 4.94万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1997
- 资助国家:美国
- 起止时间:1997-03-01 至 2011-02-28
- 项目状态:已结题
- 来源:
- 关键词:AnimalsBiological AssayCell physiologyCellsCleaved cellCountDNADNA DamageDefectDependovirusDetectionDiseaseDouble Strand Break RepairEffectivenessEventExonsEye diseasesFluorescenceFoundationsFrequenciesGene ExpressionGenesGenomeGenomicsGoalsGrantGreen Fluorescent ProteinsHumanIndividualIntronsKnock-in MouseKnock-outLeadLocalizedMeasuresMessenger RNAMetabolismMethodsMusMutateMutationNeurodegenerative DisordersNeuronsNonsense-Mediated DecayOcular PhysiologyPhotoreceptorsPoisonProceduresProcessPropertyProteinsRangeReagentResearch PersonnelRetinaRetinal DegenerationRetinitis PigmentosaRhodopsinRod Outer SegmentsSiteStructureSystemTestingTherapeuticZinc Fingersadeno-associated viral vectorbasedesignfusion genegene correctiongene repairgenetic manipulationhomologous recombinationknockout genemouse genomemutantnovelnucleaseprogramsrepairedresearch studyretinal rodssubretinal injectiontherapy design
项目摘要
Strategies for correction and knockout of the rhodopsin gene in rod photoreceptor cells will be tested to
determine what it takes to manipulate the structure and expression of deleterious genes in terminally
differentiated neurons. We seek to understand the fundamental cellular processes that allow targeted gene
repair and mutation in these cells, as well as those that can control expression of toxic proteins. In the process
of gaining this understanding, we aim to develop effective methods for treating autosomal dominant retinitis
pigmentosa (ADRP) caused by defects in the rhodopsin gene. Successful therapies for ADRP will provide a
paradigm for treatment of dominant eye diseases and other neurodegenerative disorders. Mice are a natural
choice for these studies because their eye physiology resembles that of humans;ADRP disease genes cause
retinal degeneration in mice more quickly than in humans and large animals, allowing rapid testing of
efficacy; and mouse genomes can be readily manipulated. Previously, we fused the complete human
rhodopsin gene¿the ultimate target for therapy¿tothe GFP gene to generate a visible marker for rhodopsin
expression that localizes properly to rod outer segments. Here, we propose to use our existing wild type
rhodopsin-GTP mice and generate four new mouse lines carrying mutant human rhodopsin-GFP genes to
provide targets for various kinds of genetic manipulation. These modified human rhodopsin-GFP genes will
include three defined ADRP mutations¿P23H, Q64ter, and Q344ter¿and a gene that carries an internal
duplication. Adeno-associated virus vectors will be used to deliver novel reagents for rhodopsin correction
and knockout. For gene correction, we will test segments of rhodopsin DNA in the presence and absence of
genes for zinc-finger nucleases (ZFNs) that have been rationally designedto cleave near the target mutations.
For gene knockout, we will test exon-specific ZFNs to mutate the rhodopsin-GFP gene directly, and intron-
specific ZFNs designed to stimulate incorporation of a 'killer' exon that will poison expression of the
rhodopsin gene. Assays for rhodopsin gene correction and knockout will utilize the fluorescent properties of
the humanrhodopsin-GFPtarget gene.
矫正和敲除视杆细胞视紫红质基因的策略将被测试,
确定需要什么来操纵有害基因的结构和表达,
分化的神经元我们试图了解基本的细胞过程,使靶基因
这些细胞中的修复和突变,以及那些可以控制有毒蛋白质表达的细胞。过程中
为了获得这种理解,我们的目标是开发治疗常染色体显性视网膜炎的有效方法,
色素沉着症(ADRP)由视紫红质基因缺陷引起。ADRP的成功疗法将提供
治疗显性眼病和其他神经退行性疾病的范例。老鼠是天生的
这些研究的选择,因为他们的眼睛生理学类似于人类;ADRP疾病基因导致
小鼠的视网膜变性比人类和大型动物更快,可以快速检测
功效;并且小鼠基因组可以容易地操纵。之前我们融合了完整的人类
视紫红质基因作为治疗的最终靶点,与绿色荧光蛋白基因结合,产生视紫红质的可见标记物
表达,适当地定位到杆外段。在这里,我们建议使用我们现有的野生型
rhodopsin-GTP小鼠,并产生四种携带突变的人rhodopsin-GFP基因的新小鼠品系,
为各种基因操作提供了目标。这些修饰的人类视紫红质-GFP基因将
包括三个确定的ADRP突变<$P23 H,Q64 ter和Q344 ter <$和一个携带内部
重复。腺相关病毒载体将被用于递送用于视紫红质校正的新型试剂
和击倒。对于基因校正,我们将在存在和不存在以下物质的情况下测试视紫红质DNA片段:
锌指核酸酶(ZFN)的基因已经被合理地设计成在靶突变附近切割。
对于基因敲除,我们将测试外显子特异性ZFN以直接突变视紫红质-GFP基因,并测试内含子-GFP基因。
特异性ZFN被设计为刺激“杀手”外显子的掺入,所述“杀手”外显子将毒害
视紫红质基因用于视紫红质基因校正和敲除的测定将利用以下的荧光性质:
人视紫红质-GFP靶基因。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
JOHN H WILSON其他文献
JOHN H WILSON的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('JOHN H WILSON', 18)}}的其他基金
Instability of Triplet Repeats in Mammalian Cells
哺乳动物细胞中三联体重复的不稳定性
- 批准号:
7904472 - 财政年份:2009
- 资助金额:
$ 4.94万 - 项目类别:
Rhodopsin Gene Correction and Gene Knockout in Rod Cells
视杆细胞中的视紫红质基因校正和基因敲除
- 批准号:
8655854 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
RHODOSPIN GENE CORRECTION BY OLIGONUCLEOTIDE TARGETING
通过寡核苷酸靶向进行视紫红质基因校正
- 批准号:
6314816 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
Rhodopsin Gene Correction and Gene Knockout in Rod Cells
视杆细胞中的视紫红质基因校正和基因敲除
- 批准号:
7186678 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
Rhodopsin Gene Correction and Gene Knockout in Rod Cells
视杆细胞中的视紫红质基因校正和基因敲除
- 批准号:
7386597 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
RHODOPSIN GENE CORRECTION BY OLIGONUCLEOTIDE TARGETING
通过寡核苷酸靶向进行视紫红质基因校正
- 批准号:
2020240 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
RHODOSPIN GENE CORRECTION BY OLIGONUCLEOTIDE TARGETING
通过寡核苷酸靶向进行视紫红质基因校正
- 批准号:
6041352 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
RHODOSPIN GENE CORRECTION BY OLIGONUCLEOTIDE TARGETING
通过寡核苷酸靶向进行视紫红质基因校正
- 批准号:
6635654 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
Rhodopsin Gene Correction and Gene Knockout in Rod Cells
视杆细胞中的视紫红质基因校正和基因敲除
- 批准号:
8260502 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
Rhodopsin Gene Correction and Gene Knockout in Rod Cells
视杆细胞中的视紫红质基因校正和基因敲除
- 批准号:
8117905 - 财政年份:1997
- 资助金额:
$ 4.94万 - 项目类别:
相似海外基金
Establishment of a new biological assay using Hydra nematocyst deployment
利用水螅刺丝囊部署建立新的生物测定方法
- 批准号:
520728-2017 - 财政年份:2017
- 资助金额:
$ 4.94万 - 项目类别:
University Undergraduate Student Research Awards
POINT-OF-CARE BIOLOGICAL ASSAY FOR DETERMINING TISSUE-SPECIFIC ABSORBED IONIZING RADIATION DOSE (BIODOSIMETER) AFTER RADIOLOGICAL AND NUCLEAR EVENTS.
用于确定放射和核事件后组织特异性吸收电离辐射剂量(生物剂量计)的护理点生物测定。
- 批准号:
10368760 - 财政年份:2017
- 资助金额:
$ 4.94万 - 项目类别:
POINT-OF-CARE BIOLOGICAL ASSAY FOR DETERMINING TISSUE-SPECIFIC ABSORBED IONIZING RADIATION DOSE (BIODOSIMETER) AFTER RADIOLOGICAL AND NUCLEAR EVENTS.
用于确定放射和核事件后组织特异性吸收电离辐射剂量(生物剂量计)的护理点生物测定。
- 批准号:
10669539 - 财政年份:2017
- 资助金额:
$ 4.94万 - 项目类别:
POINT-OF-CARE BIOLOGICAL ASSAY FOR DETERMINING TISSUE-SPECIFIC ABSORBED IONIZING RADIATION DOSE (BIODOSIMETER) AFTER RADIOLOGICAL AND NUCLEAR EVENTS.
用于确定放射和核事件后组织特异性吸收电离辐射剂量(生物剂量计)的护理点生物测定。
- 批准号:
9570142 - 财政年份:2017
- 资助金额:
$ 4.94万 - 项目类别:
POINT-OF-CARE BIOLOGICAL ASSAY FOR DETERMINING TISSUE-SPECIFIC ABSORBED IONIZING RADIATION DOSE (BIODOSIMETER) AFTER RADIOLOGICAL AND NUCLEAR EVENTS.
用于确定放射和核事件后组织特异性吸收电离辐射剂量(生物剂量计)的护理点生物测定。
- 批准号:
9915803 - 财政年份:2017
- 资助金额:
$ 4.94万 - 项目类别:
COVID-19 Supplemental work: POINT-OF-CARE BIOLOGICAL ASSAY FOR DETERMINING TISSUE-SPECIFIC ABSORBED IONIZING RADIATION DOSE (BIODOSIMETER).
COVID-19 补充工作:用于确定组织特异性吸收电离辐射剂量的护理点生物测定(生物剂量计)。
- 批准号:
10259999 - 财政年份:2017
- 资助金额:
$ 4.94万 - 项目类别:
Drug discovery based on a new biological assay system using Yeast knock-out strain collection
基于使用酵母敲除菌株收集的新生物测定系统的药物发现
- 批准号:
21580130 - 财政年份:2009
- 资助金额:
$ 4.94万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Machine learning for automatic gene annotation using high-throughput biological assay data
使用高通量生物测定数据进行自动基因注释的机器学习
- 批准号:
300985-2004 - 财政年份:2005
- 资助金额:
$ 4.94万 - 项目类别:
Postdoctoral Fellowships
Machine learning for automatic gene annotation using high-throughput biological assay data
使用高通量生物测定数据进行自动基因注释的机器学习
- 批准号:
300985-2004 - 财政年份:2004
- 资助金额:
$ 4.94万 - 项目类别:
Postdoctoral Fellowships