Introduction to my KCGS CDK family project

At the SGC-UNC we are interested in building a set of inhibitors that “covers” the whole kinome. Here’s a reference that talks about our plans for the Kinase Chemogenomic Set (KCGS): http://journals.plos.org/plosone/article/related?id=10.1371/journal.pone.0181585. We are building the set with what we call “narrow spectrum” inhibitors – inhibitors that inhibit only a small set of kinases. To help Read More …

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 …

Western Blot of NSD3 in AML Cell Lines

In order to study NSD3 in the context of acute myeloid leukemia, I first need to determine the best reagents/tools. This includes characterizing model cell lines as well as antibodies that can be used for future experiments. Here I looked at the expression of NSD3 by western blotting in three AML cell lines using a 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 …