The Role Of Nestin Cre Recombinase Expression Patterns in E15.5 Transgenic Mice Embryos and Neuronal Stem Cell Lines In Recurrent DNA Break Clusters (RDCs)
School Name
South Carolina Governor's School for Science and Mathematics
Grade Level
12th Grade
Presentation Topic
Cell and Molecular Biology
Presentation Type
Mentored
Abstract
Recurrent DNA Break Clusters (RDCs) are double stranded breaks that occur within the gene body of cell adhesion molecules (CAM). RDCs are highly prevalent in neural stem and progenitor cells and have been shown to cause neuropsychiatric disorders and cancer, and occur more frequently in the presence of stressors. This research aimed to study two stressors: cell proliferation and the addition of aphidicolin [DNA polymerase inhibitor] to embryonic stem cells. In order to observe the first stressor, transgenic mice embryos in the E15.5 stage containing the TdT reporter gene as well as the Nestin-Cre Protein expression vectors, were used. The transgenic mice embryos contained the LoxP Stop LoxP (LSP) sequence that affected the expression of the TdTomato protein in the transgenic mice embryos. The morphology of Nestin Cre (+/-) embryos were sliced and stained using the following staining methods: Hematoxylin and Eosin (H&E) to observe general morphology; Nissl to observe pathology; and Immunofluorescence to observe TdTomato presence in the organism. The results indicate that the presence of Nestin-Cre led to TdTomato expression, and normal pathology within the embryos. In addition, pluripotent neural stem cells were generated using the Leukemia Inhibitory Factors (LIF) in the NXP010 [p53- and XRCC4-] embryonic stem cells. These cells are very susceptible to double stranded breaks, and therefore were used in this research to better observe RDCs in-vitro and systemically.
Recommended Citation
Koruk, Sare, "The Role Of Nestin Cre Recombinase Expression Patterns in E15.5 Transgenic Mice Embryos and Neuronal Stem Cell Lines In Recurrent DNA Break Clusters (RDCs)" (2023). South Carolina Junior Academy of Science. 14.
https://scholarexchange.furman.edu/scjas/2023/all/14
Location
ECL 104
Start Date
3-25-2023 11:00 AM
Presentation Format
Oral Only
Group Project
No
The Role Of Nestin Cre Recombinase Expression Patterns in E15.5 Transgenic Mice Embryos and Neuronal Stem Cell Lines In Recurrent DNA Break Clusters (RDCs)
ECL 104
Recurrent DNA Break Clusters (RDCs) are double stranded breaks that occur within the gene body of cell adhesion molecules (CAM). RDCs are highly prevalent in neural stem and progenitor cells and have been shown to cause neuropsychiatric disorders and cancer, and occur more frequently in the presence of stressors. This research aimed to study two stressors: cell proliferation and the addition of aphidicolin [DNA polymerase inhibitor] to embryonic stem cells. In order to observe the first stressor, transgenic mice embryos in the E15.5 stage containing the TdT reporter gene as well as the Nestin-Cre Protein expression vectors, were used. The transgenic mice embryos contained the LoxP Stop LoxP (LSP) sequence that affected the expression of the TdTomato protein in the transgenic mice embryos. The morphology of Nestin Cre (+/-) embryos were sliced and stained using the following staining methods: Hematoxylin and Eosin (H&E) to observe general morphology; Nissl to observe pathology; and Immunofluorescence to observe TdTomato presence in the organism. The results indicate that the presence of Nestin-Cre led to TdTomato expression, and normal pathology within the embryos. In addition, pluripotent neural stem cells were generated using the Leukemia Inhibitory Factors (LIF) in the NXP010 [p53- and XRCC4-] embryonic stem cells. These cells are very susceptible to double stranded breaks, and therefore were used in this research to better observe RDCs in-vitro and systemically.
