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Fluorescence stereomicroscope imaging systems

荧光立体显微镜成像系统

基本信息

批准号：
10601232
负责人：
Jeffrey J Essner
金额：
$ 25.85万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2024-03-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10601232
关键词：
Administrative Supplement Alleles Blood Vessels Cell Lineage Cells Clustered Regularly Interspaced Short Palindromic Repeats Communities Complex Computer software Computers Cre driver Data Detection Development Disease Documentation Embryo Engineered Gene Ensure Enterobacteria phage P1 Cre recombinase Equipment Failure Fluorescence Fluorescence Microscopy Funding Future Gene Expression Genes Goals Grant Health Heart Hematopoietic Human Human Development Image Immune Individual Injury Knock-out Label Larva LoxP-flanked allele Methods Microscope Monoclonal Antibody R24 Organ Parents Process Reporter Reproducibility Research Resolution Resources Specific qualifier value System Time Tissues Transgenic Organisms Vertebrates Visualization Zebrafish cell type developmental disease gene cloning gene function genome editing human disease human model imaging capabilities imaging system improved in vivo in vivo fluorescence innovation insight interest knockout gene live cell imaging novel novel strategies programs promoter real-time images routine screening screening tool transcription factor zebrafish genome

项目摘要

Fluorescence stereomicroscopy for live imaging of zebrafish embryos and larvae has revolutionized our ability to gain insight into human developmental and disease processes. It allows visualization in real time of cell lineages that build the complex tissues and organs that define vertebrate animals. Examining how these processes are disrupted by injury or disease leads to novel discoveries of the cellular mechanisms that can be manipulated to improve human health. The vast amount of single cell expression data generated from distinct developmental stages and tissues has provided a wealth of new information about key transcription factors that specify cell lineages. Until now, methods to create transcription factor-specific cell lineage tracing reporters, such as Cre recombinase, have been inefficient and limited by the ability to clone gene specific promoters. With our GeneWeld CRISPR precision targeted integration strategy, we can generate a Cre driver in any gene of interest, unlocking the potential to examine cell lineages that were previously inaccessible, and to transform single cell gene expression data into functional tools for cell lineage specific analysis. With our R24 resource grant, we have recovered Cre drivers in transcription factor genes that define mesodermal and immune cell lineages and are expanding this to early mesendoderm, vascular, and hematopoietic cells. Fluorescence microscopy is at the heart of the transgenic resource we are building and functional fluorescence stereomicroscope equipment is critical to efficiently generate and characterize new lines. To meet the imaging needs of the R24, we propose 2 aims that will significantly expand our fluorescence stereomicroscope capabilities. In Aim 1 we propose funds to support our imaging capabilities through the purchase of 3 Leica M165 FC imaging systems for routine daily screening and documentation of new transgenic lines. In Aim 2 we propose funds to elevate our capacity for real time live imaging of cell lineage labeling through the purchase of 1 Leica M205 FCA Thunder imaging system. Together, these systems will allow us to successfully grow the zebrafish Cre driver community resource and thoroughly characterize the spatial and temporal cell lineage labeling activity of each Cre line.

荧光立体显微镜用于斑马鱼胚胎和幼虫的实时成像已经彻底改变了我们的

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Endogenous zebrafish proneural Cre drivers generated by CRISPR/Cas9 short homology directed targeted integration.

DOI：
10.1038/s41598-021-81239-y
发表时间：
2021-01-18
期刊：
Scientific reports
影响因子：
4.6
作者：
Almeida MP;Welker JM;Siddiqui S;Luiken J;Ekker SC;Clark KJ;Essner JJ;McGrail M
通讯作者：
McGrail M

Cre/lox regulated conditional rescue and inactivation with zebrafish UFlip alleles generated by CRISPR-Cas9 targeted integration.

DOI：
10.7554/elife.71478
发表时间：
2022-06-17
期刊：
ELIFE
影响因子：
7.7
作者：
Liu, Fang;Kambakam, Sekhar;Almeida, Maira P.;Ming, Zhitao;Welker, Jordan M.;Wierson, Wesley A.;Schultz-Rogers, Laura E.;Ekker, Stephen C.;Clark, Karl J.;Essner, Jeffrey J.;McGrail, Maura
通讯作者：
McGrail, Maura

Epigenetic regulators Rbbp4 and Hdac1 are overexpressed in a zebrafish model of RB1 embryonal brain tumor, and are required for neural progenitor survival and proliferation.

DOI：
10.1242/dmm.034124
发表时间：
2018-06-15
期刊：
Disease models & mechanisms
影响因子：
4.3
作者：
Schultz LE;Haltom JA;Almeida MP;Wierson WA;Solin SL;Weiss TJ;Helmer JA;Sandquist EJ;Shive HR;McGrail M
通讯作者：
McGrail M