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Cell line Core

细胞系核心

基本信息

批准号：
10202526
负责人：
Gaorav P Gupta
金额：
$ 12.48万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
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：
2020-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10202526
关键词：
Aliquot Alleles Biological Biological Assay CRISPR/Cas technology Cell Line Cells Clone Cells Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Communities Credentialing Cryopreservation Custom DNA Double Strand Break DNA Repair Enzymes DNA-Directed DNA Polymerase Doxycycline Engineering Ensure Enzymes Experimental Designs Future Gene Targeting Generations Genes Genetic Engineering Genome Genome engineering Genomic Instability Genomics Genotype Goals Human In Vitro Karyotype Karyotype determination procedure Knock-in Knock-out Ligands Malignant Neoplasms Mammalian Cell Manuscripts Mediating Methods Modeling Monitor Mus Mycoplasma Polymerase Process Proteins Protocols documentation Publications Quality Control Reagent Reproducibility Research Research Activity Research Personnel Role Secure Services Standardization Structure of thyroid parafollicular cell System Testing Time Transgenes Validation base cost effectiveness design digital empowered experimental study genetically modified cells homologous recombination in vivo interest laboratory experiment multidisciplinary overexpression parity programs protein expression quality assurance repaired repository screening service programs success synergism transcriptome sequencing transgene delivery transgene expression vector

项目摘要

PROJECT SUMMARY This project will investigate mammalian DNA polymerase θ (Pol θ), the defining enzyme for repair of DNA double-strand breaks by polymerase theta-mediated end joining (TMEJ). This is Core C (“Cell Line Engineering Core”) which is part of a Program Project titled, “Polymerase theta, genome instability, and cancer”. Core C will be responsible for providing native and genetically engineered cell lines to support all of the cell-based assays performed in Projects 1, 2, and 4 of the Program Project. The authentication of mammalian cell line reagents is vital for rigor and reproducibility of laboratory experiments. By establishing and implementing standardized protocols for cell line validation, efficient genome manipulation, cryopreservation, and dissemination, the Core will ensure quality control of cell line reagents and minimize the likelihood of erroneous observations and interpretations. In Aim 1, “Credentialing of mammalian cell line models utilized in this Program Project”, we will implement a standardized and comprehensive protocol for authentication and characterization of mammalian cell line models used across the Program Project. In Aim 2, “CRISPR-based cell line genome engineering services for the Program Project”, we will provide expert services in the design and execution of customized cell line genome engineering tasks in support of Program Project goals. Cas9 protein utilized in this Aim will be provided by Core B. In Aim 3, “Controlled and stable expression of transgenes in mammalian cell lines”, we will utilize doxycycline-inducible and constitutive transposon vectors for the generation of cell lines that express a variety of mutagenized and/or tagged transgenes of interest. Core C services will be highly coordinated within the Program Project, and will facilitate rigor and reproducibility, as well as cost-effectiveness, of highly integrated research activities aimed at understanding the role and targetability of DNA Polymerase q in cancer.

项目总结