Towards a genome-wide CRISPR/Cas9 mutant library in Rhizopus delemar
德莱马根霉 (Rhizopus delemar) 中的全基因组 CRISPR/Cas9 突变体文库
基本信息
- 批准号:10573271
- 负责人:
- 金额:$ 18.38万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-02-15 至 2025-01-31
- 项目状态:未结题
- 来源:
- 关键词:Amphotericin BCRISPR/Cas technologyClinicalCodeCollectionConsumptionDNA Polymerase IIIDNA sequencingDideoxy Chain Termination DNA SequencingFungal Drug ResistanceFutureGene MutationGenerationsGenesGeneticGenomeGenotypeGlycine-Specific tRNAGrowthGuide RNAHealthHumanImmunologic Deficiency SyndromesInduced MutationLibrariesLifeLigationMolecularMolecular DiseaseMorphologyMucormycosisMutagenesisMutationNutritional RequirementsPathogenesisPatientsPhenotypePoint MutationProteinsRNARandom AllocationResearchResearch Project GrantsResistanceResourcesRhizopusSequence HomologySpecificitySystemTimeVirulencedesignefficacy evaluationexpression vectorgene gungenome-wideimprovedinnovationmortalitymutantnext generation sequencingoff-target mutationpathogenic funguspromotersuccesstoolvectorwhole genome
项目摘要
Rhizopus delemar is the primary cause of life-threatening mucormycosis
responsible for
approximately 70% of clinical cases, with an overall mortality rate exceeding 50%. Despite its
devastating impact on human health,
much of R. delemar’s pathobiology and molecular disease
mechanisms remains unknown. As an initial step towards constructing a high-throughput genome-wide
mutant library of R. delemar, we propose
to generate a small-scale targeted mutant library utilizing
multiplex CRISPR/Cas9 technology. In Specific Aim 1, we will synthesize a collection of single-guide
RNAs (sgRNAs) targeting 40 randomly selected genes and deliver pooled sgRNAs and Cas9 in a one-
vector construct into R. delemar. In Specific Aim 2, we will identify gene mutations using Sanger DNA
sequencing and high-throughput next-generation sequencing. We will also assess efficacy, specificity,
and off-target mutations induced by multiplex CRISPR/Cas9. The success of this exploratory research
effort will lead to the construction of whole-genome CRISPR/Cas9 mutant libraries for the systemic
examination of R. delemar pathogenesis mechanisms.
德尔玛根霉是威胁生命的毛霉病的主要原因
负责
约70%的临床病例,总死亡率超过50%。尽管它的
对人类健康造成毁灭性的影响,
德尔马氏杆菌的大部分病理生物学和分子疾病
机制仍不清楚。作为构建高通量全基因组
的突变体文库,我们建议
利用以下技术生成小规模靶向突变体文库
多路复用CRISPR/CAS9技术。在具体目标1中,我们将综合一组单一指南
针对40个随机选择的基因的RNA(SgRNAs),并将汇集的sgRNAs和Cas9合二为一-
将载体构建到R.delemar。在特定目标2中,我们将使用Sanger DNA识别基因突变
测序和高通量下一代测序。我们还将评估疗效、特异性、
和多重CRISPR/Cas9诱发的靶外突变。这项探索性研究的成功
这项工作将导致构建全基因组CRISPR/Cas9突变文库,用于系统
德尔玛黄粉菌致病机理的研究。
项目成果
期刊论文数量(12)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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- 发表时间:2022-09-15
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- DOI:10.7554/elife.85728
- 发表时间:2024-01-22
- 期刊:
- 影响因子:7.7
- 作者:Meng Y;Ni Y;Li Z;Jiang T;Sun T;Li Y;Gao X;Li H;Suo C;Li C;Yang S;Lan T;Liao G;Liu T;Wang P;Ding C
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- DOI:10.1111/mpp.13225
- 发表时间:2022-08
- 期刊:
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- DOI:10.1371/journal.ppat.1011251
- 发表时间:2023-04
- 期刊:
- 影响因子:6.7
- 作者:
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{{ truncateString('PING WANG', 18)}}的其他基金
Towards a genome-wide CRISPR/Cas9 mutant library in Rhizopus delemar
德莱马根霉 (Rhizopus delemar) 中的全基因组 CRISPR/Cas9 突变体文库
- 批准号:
10431481 - 财政年份:2022
- 资助金额:
$ 18.38万 - 项目类别:
Isolation of mononuclear propagules from coenocytic hyphae of the mucormycosis pathogen Rhizopus delemar
从毛霉菌病病原体德莱马根霉的共生菌丝中分离单核繁殖体
- 批准号:
10353429 - 财政年份:2021
- 资助金额:
$ 18.38万 - 项目类别:
Isolation of mononuclear propagules from coenocytic hyphae of the mucormycosis pathogen Rhizopus delemar
从毛霉菌病病原体德莱马根霉的共生菌丝中分离单核繁殖体
- 批准号:
10221180 - 财政年份:2021
- 资助金额:
$ 18.38万 - 项目类别:
RADX TECH PROJECT - WORK PACKAGE 1 SUPPORT
RADX 技术项目 - 工作包 1 支持
- 批准号:
10505995 - 财政年份:2021
- 资助金额:
$ 18.38万 - 项目类别:
Mechanisms of Radiation-induced Vascular Endothelial Cell Injury and Its Correction
辐射引起的血管内皮细胞损伤的机制及其纠正
- 批准号:
9391119 - 财政年份:2017
- 资助金额:
$ 18.38万 - 项目类别:
Genome editing in Rhizopus delemar using using CRISPR-Cas systems
使用 CRISPR-Cas 系统对德勒马根霉进行基因组编辑
- 批准号:
9179413 - 财政年份:2016
- 资助金额:
$ 18.38万 - 项目类别:
Novel Approaches to Maintaining Organ Function in Sepsis
维持脓毒症器官功能的新方法
- 批准号:
10405950 - 财政年份:2016
- 资助金额:
$ 18.38万 - 项目类别:
Novel Approaches to Maintaining Organ Function in Sepsis
维持脓毒症器官功能的新方法
- 批准号:
10153818 - 财政年份:2016
- 资助金额:
$ 18.38万 - 项目类别:
Genome editing in Rhizopus delemar using using CRISPR-Cas systems
使用 CRISPR-Cas 系统对德勒马根霉进行基因组编辑
- 批准号:
9304959 - 财政年份:2016
- 资助金额:
$ 18.38万 - 项目类别:
Novel Approaches to Maintaining Organ Function in Sepsis
维持脓毒症器官功能的新方法
- 批准号:
9698963 - 财政年份:2016
- 资助金额:
$ 18.38万 - 项目类别:
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