Analysis of homolog-based CRISPR editing in somatic cells
体细胞中基于同源物的 CRISPR 编辑分析
基本信息
- 批准号:10343429
- 负责人:
- 金额:$ 31.6万
- 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:2022-08-15 至 2026-06-30
- 项目状态:未结题
- 来源:
- 关键词:AdultAffectAgricultureAllelesBacteriaBiologyCRISPR gene driveCell CycleCell LineCellsChromosome MappingChromosome PairingChromosomesClustered Regularly Interspaced Short Palindromic RepeatsCulicidaeCystic FibrosisCystic Fibrosis Transmembrane Conductance RegulatorDNADNA RepairDNA cassetteDisease modelDrosophila genusEctopic ExpressionElementsEndonuclease IEventFrequenciesG2 PhaseGene ConversionGenesGeneticGenomeGenomicsGrantGuide RNAHomologous GeneHumanHuman Cell LineInduced MutationInsect ControlInsectaMammalian CellMammalsMediatingMedicineMeiosisMitoticModelingMutationNonhomologous DNA End JoiningOncogenesOrganismOutcomePathway interactionsPhenotypePigmentation physiologic functionPopulationProcessReporterResistanceS phaseSequence HomologsSiteSomatic CellSystemTechnologyTestingVariantVisualizationYeastsbasebase editingdesigndevelopmental geneticsdisease-causing mutationexpectationexperimental studyflygastrointestinal epitheliumgene correctiongene drive allelegene drive systemgene therapygenetic elementgenetic variantgenome editinggenome integritygenomic locusimprovedin vivoinsect disease vectorinsightmutantnext generationnotch proteinnovelnovel strategiesnucleaseportabilityprecise genome editingprecursor cellrepair modelrepair strategyrepairedstem cellstooltransgene expression
项目摘要
Recently developed CRISPR-based systems permit precise genome editing by inducing targeted DNA
breaks at specific sites in the genome. Cellular DNA repair machinery can restore genome integrity by copying
information from the intact homologous chromosome at the cleavage site via homology directed repair (HDR).
While precise HDR-mediated DNA repair is the predominant pathway active during meiosis, the competing and
potentially mutagenic non-homologous end-joining pathway (NHEJ) is typically thought to prevail in somatic
cells. One reason for this bias is that the NHEJ pathway is active throughout somatic cell cycles, while HDR is
primarily restricted to post-replicative S and G2 phases. Thus, achieving efficient HDR-based gene editing in
somatic cells has proven challenging, which limits the in vivo use of this technology for human gene therapy.
My group has contributed to developing the first CRISPR-based gene-drive (or active genetic) systems in
flies, mosquitoes, mammals, and bacteria that bias germline inheritance of genetic elements programmed to
cut the genome at their site of insertion. We also pioneered allelic-drive systems designed to promote biased
inheritance of a favored allelic variant at a separate genetic locus. These germline drive systems also produce
somatic phenotypes, which have generally been attributed to mutations induced by the NHEJ pathway.
Recently, we developed genetic elements we refer to as “CopyCatchers” that permit visualization of HDR-
mediated copying of gene cassettes. These studies have revealed an unexpectedly high frequency of somatic
gene conversion (SGC) events in Drosophila (30-50%) wherein the chromosome homolog serves as a DNA
repair template. Rates of SGC can be improved further by optimizing delivery of CRISPR components, or by
reducing the expression of various genes encoding factors involved in DNA repair or chromosome pairing.
Preliminary experiments indicate that interhomolog SGC can also take place in human cells and point to
untapped strategies for repairing disease-causing mutations using intact sequences from the homologous
chromosome.
In this grant we propose to explore SGC repair mechanisms mediated by Cas9 and Nickase in somatic
cells of Drosophila and then extend analysis of this interhomolog repair process to human cells. First, we will
analyze the mechanisms underlying CRISPR dependent copying of gene cassettes or allelic variants to
optimize their activities. Next, we will develop and optimize Drosophila models for homolog-based repair of
disease-causing mutations in the Notch locus affecting mitotically active stem cells or in post-mitotic cells in the
adult gut epithelium using a humanized Drosophila CFTR–/– disease model. Finally, we will assess whether
insights gained in Drosophila are portable to human somatic cell lines, and whether interhomolog SGC can
restore native gene activity in human cell-based models for cystic fibrosis. Enhancing homolog-based repair in
mammalian cells could offer transformative possibilities for next-generation gene therapy strategies.
最近开发的基于crispr的系统可以通过诱导目标DNA进行精确的基因组编辑
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 }}
ETHAN BIER其他文献
ETHAN BIER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('ETHAN BIER', 18)}}的其他基金
Analysis of homolog-based CRISPR editing in somatic cells
体细胞中基于同源物的 CRISPR 编辑分析
- 批准号:
10676726 - 财政年份:2022
- 资助金额:
$ 31.6万 - 项目类别:
Development of next-generation gene drive technologies for Anopheles population engineering
开发用于按蚊种群工程的下一代基因驱动技术
- 批准号:
10278897 - 财政年份:2021
- 资助金额:
$ 31.6万 - 项目类别:
Development of next-generation gene drive technologies for Anopheles population engineering
开发用于按蚊种群工程的下一代基因驱动技术
- 批准号:
10624305 - 财政年份:2021
- 资助金额:
$ 31.6万 - 项目类别:
Development of next-generation gene drive technologies for Anopheles population engineering
开发用于按蚊种群工程的下一代基因驱动技术
- 批准号:
10408862 - 财政年份:2021
- 资助金额:
$ 31.6万 - 项目类别:
The mutagenic chain reaction: a method for autocatalyic gene dissemination
诱变链式反应:一种自催化基因传播的方法
- 批准号:
10211352 - 财政年份:2016
- 资助金额:
$ 31.6万 - 项目类别:
The mutagenic chain reaction: a method for autocatalyic gene dissemination
诱变链式反应:一种自催化基因传播的方法
- 批准号:
9009589 - 财政年份:2016
- 资助金额:
$ 31.6万 - 项目类别:
The mutagenic chain reaction: a method for autocatalyic gene dissemination
诱变链式反应:一种自催化基因传播的方法
- 批准号:
10395549 - 财政年份:2016
- 资助金额:
$ 31.6万 - 项目类别:
The mutagenic chain reaction: a method for autocatalyic gene dissemination
诱变链式反应:一种自催化基因传播的方法
- 批准号:
10614935 - 财政年份:2016
- 资助金额:
$ 31.6万 - 项目类别:
Mutagenic chain reaction-facilitated immunotherapy
诱变链式反应促进的免疫疗法
- 批准号:
9163059 - 财政年份:2016
- 资助金额:
$ 31.6万 - 项目类别:
Mutagenic chain reaction-facilitated immunotherapy
诱变链式反应促进的免疫疗法
- 批准号:
9755350 - 财政年份:2016
- 资助金额:
$ 31.6万 - 项目类别:
相似海外基金
How Does Particle Material Properties Insoluble and Partially Soluble Affect Sensory Perception Of Fat based Products
不溶性和部分可溶的颗粒材料特性如何影响脂肪基产品的感官知觉
- 批准号:
BB/Z514391/1 - 财政年份:2024
- 资助金额:
$ 31.6万 - 项目类别:
Training Grant
BRC-BIO: Establishing Astrangia poculata as a study system to understand how multi-partner symbiotic interactions affect pathogen response in cnidarians
BRC-BIO:建立 Astrangia poculata 作为研究系统,以了解多伙伴共生相互作用如何影响刺胞动物的病原体反应
- 批准号:
2312555 - 财政年份:2024
- 资助金额:
$ 31.6万 - 项目类别:
Standard Grant
RII Track-4:NSF: From the Ground Up to the Air Above Coastal Dunes: How Groundwater and Evaporation Affect the Mechanism of Wind Erosion
RII Track-4:NSF:从地面到沿海沙丘上方的空气:地下水和蒸发如何影响风蚀机制
- 批准号:
2327346 - 财政年份:2024
- 资助金额:
$ 31.6万 - 项目类别:
Standard Grant
Graduating in Austerity: Do Welfare Cuts Affect the Career Path of University Students?
紧缩毕业:福利削减会影响大学生的职业道路吗?
- 批准号:
ES/Z502595/1 - 财政年份:2024
- 资助金额:
$ 31.6万 - 项目类别:
Fellowship
感性個人差指標 Affect-X の構築とビスポークAIサービスの基盤確立
建立个人敏感度指数 Affect-X 并为定制人工智能服务奠定基础
- 批准号:
23K24936 - 财政年份:2024
- 资助金额:
$ 31.6万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Insecure lives and the policy disconnect: How multiple insecurities affect Levelling Up and what joined-up policy can do to help
不安全的生活和政策脱节:多种不安全因素如何影响升级以及联合政策可以提供哪些帮助
- 批准号:
ES/Z000149/1 - 财政年份:2024
- 资助金额:
$ 31.6万 - 项目类别:
Research Grant
How does metal binding affect the function of proteins targeted by a devastating pathogen of cereal crops?
金属结合如何影响谷类作物毁灭性病原体靶向的蛋白质的功能?
- 批准号:
2901648 - 财政年份:2024
- 资助金额:
$ 31.6万 - 项目类别:
Studentship
Investigating how double-negative T cells affect anti-leukemic and GvHD-inducing activities of conventional T cells
研究双阴性 T 细胞如何影响传统 T 细胞的抗白血病和 GvHD 诱导活性
- 批准号:
488039 - 财政年份:2023
- 资助金额:
$ 31.6万 - 项目类别:
Operating Grants
New Tendencies of French Film Theory: Representation, Body, Affect
法国电影理论新动向:再现、身体、情感
- 批准号:
23K00129 - 财政年份:2023
- 资助金额:
$ 31.6万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
The Protruding Void: Mystical Affect in Samuel Beckett's Prose
突出的虚空:塞缪尔·贝克特散文中的神秘影响
- 批准号:
2883985 - 财政年份:2023
- 资助金额:
$ 31.6万 - 项目类别:
Studentship














{{item.name}}会员




