I am a new member of OpenWetWare!
- Layla Barkal
- 320 Memorial Drive
- Cambridge, MA 02139
I work in the Chang lab at MIT. I learned about OpenWetWare through 20.109, a lab class I'm currently taking at MIT.
- 2011, BS, MIT
Impact on Differentiation of Ago2 Protein Lacking Slicer Functionality
Topic: Differentiation of stem cells, despite being a topic of much research, involves a variety of mechanisms, such as the miRNA pathway, that are not yet understood. In the miRNA pathway, there are various methods of regulation that have yet to be explored. To do so, we propose an experiment to target a specific element of the miRNA pathway and compare differentiation patterns with the standard hanging-drop control, thus making our work more relevant and accessible to the scientific community studying differentiation.
Project Idea: To test the effect of Ago2 with and without slicer function of the differentiation on mouse stem cells (MS cells), we propose an experiment that would directly compare at specific time intervals, the differentiation patterns of wild type MS cells, MS cells with an Ago2 protein lacking slicer functionality, with MES cells lacking the Ago2 protein all together. Analyzing levels of specific growth factors (with semi-quantitative RT-PCR) will allow us to identify the ratios in which differentiated cell types coexist. By taking RNA samples at different time points during the differentiation process, we will be able to see how the cell type ratios change over time and between the three samples (control, Ago2 lacking slicer function, and Ago2 -/-). Overall, we hope to learn more about the role Ago2 plays in differentiation of MS cells, specifically which cell types are most influenced and when.
1. A Slicer-Independent Role for Argonaut2 in Hematopoiesis and the miRNA Pathway: (here)
In this paper, the authors explain their findings that indicate that Argonaute2 has a function in the differentiation pathway that goes beyond its function as a slicing part of the RISC complex involved in miRNA regulation. They tested mice that were deficient in Ago2 and mice that had a form of Ago2 that lacked slicer function. They then analyzed the impact of these changes on hematopoiesis, looking specifically at early differentiation of bone marrow stem cells into erythroid and lymphoid cells. While they found that Ago2 was required for differentiation, slicer function was not required.
2. Novel MicroRNA Candidates and miRNA-mRNA Pairs in Embryonic Stem (ES) Cells: (here)
This paper studies the role of RNA in proliferation and differentiation of stem cells. The authors worked on Dicer -/- cells and expected the phenotypes of Dicer-/- cells to be due to problems in differentiation regulating pathways, usually mediated by miRNAs.