Optimising transfection of the C2C12 myoblast cell line

Within a month (hopefully) I’ll be convincing C2C12 myoblast cells to express all kinds of mutant ALK2 to test its activity and interactions with other proteins within the cell. I’ll be doing this by treating them so that they’ll take up pieces of circular DNA (plasmids) that express those mutant proteins (i.e. transfecting them). Seeing Read More …

Structure Activity Relationship (SAR) study to identify Nek4 inhibitors:

Hello! Recently, I co-authored a manuscript titled, “In depth analysis of kinase cross screening data to identify chemical starting points for inhibition of the Nek family of kinases” [http://pubs.rsc.org/en/content/articlelanding/2018/md/c7md00510e#!divAbstract]. As a part of this manuscript we identified several chemical starting points that can be used for the design of narrow spectrum and potent Nek kinase Read More …

Analyzing the structural diversity of the USP5 Zf-UBD binding pocket

USP5 is one of many USP proteins that contains a zinc finger ubiquitin binding domain (Zf-UBD). My goal is to develop small molecules that inhibit USP5 selectively. So, it’s important to make sure that the pocket where the compound will bind is sufficiently and structurally different from other USPs. I did a multiple sequence alignment Read More …

Project overview: Establishing Cellular Assays to Screen for ALK2 Inhibitors

Diffuse Intrinsic Pontine Glioma (DIPG) is a type of brain tumour in the brainstem which is highly aggressive and occurs in children. Treatment options for DIPG are very limited because these tumours do not respond to the chemotherapy drugs currently available for adult gliomas. Whole genome and exome sequencing identified several frequently mutated genes in Read More …

ACVR1 – the link between FOP and DIPG

Background What is the link between a rare genetic disease that causes soft tissues to turn to bone, and a lethal childhood brain cancer? At first glance, with such different clinical phenotypes, it seems unlikely there could be any link, but it has been shown through whole genome sequencing of diffuse intrinsic pontine glioma (DIPG) Read More …

Cloning an NSD3-Short-3xFLAG Construct into a Lentiviral Expression System

Antibodies are fundamental tools in the exploration of cellular biology. Unfortunately, there is a lack of high-quality NSD3 antibodies available. To overcome this, I have generated a bicistronic NSD3-Short-3xFLAG – IRES – PuroR lentiviral expression plasmid for the purpose of creating cell lines that stably express 3xFLAG-tagged NSD3-short. This will allow me to use a Read More …

Generation of fluorescently labelled OP9 stromal cells for growing patient leukemic cells

Our experiments use cells from patients with leukemia. These primary cells are notoriously difficult to culture, especially long term. To overcome this, we are using a co-culture system where we grow the leukemia cells with mouse stromal cells (OP9) to try to mimic some aspects of the bone marrow environment present in leukemia. Although essential Read More …

Chemical probes for understudied kinases

One of the ongoing projects at SGC-UNC is the development of chemical probes for understudied kinases (e.g. MST1-4, TAOK1-3, DCAMKL1, MAP3K2 and MAP3K3). A chemical probe is defined as a small molecule that selectively and potently modulates a protein’s function. Generating chemical probes for understudied kinases is important so we can further understand the biology Read More …