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Synthetic recording and in situ readout of cell lineage and molecular history in mammalian retina

哺乳动物视网膜细胞谱系和分子历史的综合记录和原位读出

基本信息

批准号：
10558663
负责人：
Amjad Askary
金额：
$ 24.9万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-30 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10558663
关键词：
Address Affect Area Award Bar Codes Biology Biomedical Engineering Birth Blindness Brain Cell Lineage Cells Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Competence Core Facility Development Developmental Biology Disease Engineering Epigenetic Process Future Gene Expression Profile Gene Expression Profiling Generations Genome Genomics Goals Grant Heterogeneity Image In Situ Individual Inherited International Intrinsic factor Knowledge Label Lead Lentivirus Vector Libraries Light Maps Measurement Mentors Mentorship Methods Molecular Mus Mutation Neuroglia Neurons Organism Pathway interactions Pattern Pilot Projects Process Readability Recording of previous events Regenerative Medicine Regulation Research Resources Retina Retinal Degeneration Role Running Signal Transduction Site Stereotyping Structure Students System Technology Time Tissues Training Training Programs Trees Viral Viral Genome Vision Visual Work Writing base editor career cell fate specification cell type cellular development design epigenetic regulation experimental study genome editing graduate student in situ imaging insight meetings member neurogenesis new technology novel organizational structure preservation progenitor programs reconstruction retinal neuron retinal progenitor cell retinal regeneration skills success synthetic biology visual information visual processing

项目摘要

Abstract The long term goal of this project is to describe how the intricate structure of mammalian retina is made during development. To achieve this goal, this proposal focuses on developing novel molecular recording technologies that enable reconstruction of cell lineage and molecular history based on in situ endpoint measurements. These methods leverage recent advances in genome editing technology to make targeted mutations in synthetic barcode arrays that make a permanent and inheritable record of history of individual cells in their genome. Importantly, the methods developed here are compatible with imaging based readout of information and can be combined with in situ transcriptional profiling to provide a unified view of the current state, past history, and spatial context of individual cells. By applying these methods, I will map the clonal structure of the mouse retina, investigate the extent to which local signals affect cell fate decisions, reveal the stochastic versus pre-programmed aspects of retinal lineage, and reconstruct the single cell trajectory of retinal progenitor cells as they proceed through different competence states. Together, these experiments provide fundamental new insight into cell fate specification process in mammalian retina and, therefore, will aid the future efforts in regenerative medicine to remedy conditions in which misspecification or degeneration of retinal neurons lead to blindness. My career goal is to run an academic lab that studies how cells acquire and maintain different identities during development, using molecular recording and synthetic biology approaches. The proposed experiments will provide me with further training in retina biology, multiplexed sequential FISH technology, and single cell genomics that are required for this goal. I have developed a detailed training plan with my mentor, Dr. Michael Elowitz, to help me transition to independence. I will meet regularly with Dr. Elowitz to discuss research progress, strategies for grant writing, student mentorship and lab management. To practice my mentorship skills, I will oversee the work of a graduate student and a technician in the lab. To broaden my scientific network, I will present my work at 2-3 international meetings per year. To seek out additional mentorship, in areas critical to the success of this project, I will collaborate with and receive guidance from Dr. Connie Cepko, expert in retina biology, Dr. Long Cai, pioneer in sequential FISH methods, and Dr. Jay Shendure, expert in single cell genomics. As a member of Caltech Division of Biology and Biological Engineering, I will have access to leaders in synthetic as well as developmental biology, cutting-edge courses and training programs, and state-of-the-art core facilities. The Pathway to Independence Award will provide the time and resources required to initiate an ambitious research program on the reconstruction of developmental history of mammalian retina.

摘要