Authors:

Emily Zhao

Date Created:

Not specified

Course Title:

Professor:

Not specified

About Paper:

YTHDF are a family of cytosolic reader proteins of the mRNA modification N6-methyladenosine (m6A). Though previously believed to hold distinct functions, the three members of the YTHDF protein family (YTHDF1, YTHDF2, YTHDF3) have been recently shown to redundantly promote degradation of their target mRNA, and exhibit compensatory responses to depletions of each other depending on cell type and levels of expression. Acute Myeloid Leukemia is characterized by the abnormal proliferation of immature myeloblasts, which has been shown to be inhibited by YTHDF protein depletion through accelerated cell death or differentiation. However, the specific mechanisms of this association remain unknown. This project stands on my lab's previous findings of YTHDF2 sufficiency for cell viability in HeLa cells under YTHDF1/3 knockout conditions, and aims to further explore and verify the findings of a previous CRISPR screening on five crucial residues identified to have yielded the greatest change in cell proliferation scores. We hypothesize the difference in cell proliferation to be caused by phosphorylation-related cellular localization of the YTHDF protein, and therefore expect to observe decreased cell survival in three of the specified sites, and aim to gather further data on the two other sites that have previously increased cell proliferation. This study employs CRISPR guided base editing administered through lentiviral transduction, and the established cell lines were then tested on a cell proliferation assay and their genome extracted for sequencing to verify mutation. However, there has not yet been sufficient data to conclusively support or deny our hypothesis.

Source:

Columbia / Biology / 2028

Topics:

No topics listed

Co-authors:

Emily Zhao