A growing team of groundbreaking scientists around the world are now sharing their lab notebooks online
The NSD protein family has been implicated in cancers, Therefore they are considered as potential targets for cancer drug discovery. Here is the Expression and purification protocol of recombinant BIO NSD3 PWWP1 for further research. https://zenodo.org/record/4088451#.X4cd6WhKi70
Background NSD3 is a member of the NSD (nuclear receptor SET domain- containing) family of H3K36 methyltransferases. It has two isoforms: long -1437 amino acid and short – 645 amino acids, which are derived from alternative splicing of exon 10. In contrast to long isoform, the short isoform lacks the methyltransferase domain and it contains Read More …
There are two major NSD3 isoforms expressed, long (aa 1-1437) and short (aa 1-645, differing in sequence from 620-645). The short isoform lacks a methyltransferase domain and, perhaps surprisingly, is the form attributed to NSD3’s role in acute myeloid leukemia (AML) [1]. I’ve previously used the TCGA-LUSC data-set [2] to test for differential expression based on the Read More …
These days CRISPR gene editing needs little introduction. From humble origins as a bacterial defence system evolved to thwart viral invaders, CRISPR-based technologies have recently made headlines for their promise to revolutionize medicine. While there is a lot of excitement surrounding the use of gene editing in the clinic, it is also an incredibly valuable tool for studying the Read More …
NSD3 is located on a region of chromosome 8 (8p11-12) that is frequently amplified in several forms of cancer. It has been suggested that NSD3 (WHSC1L1) may be a driver of tumorigenesis in this context (Mahmood et al. 2013). However, it is still unclear how amplification of NSD3 contributes to the disease and whether or not it Read More …
To understand what proteins do, scientists often test the consequence their absence has on a cell. A common method uses a cells regulatory machinery to disrupt gene expression with complementary RNA molecules, known as RNA interference. Here I used small interfering RNAs (siRNAs) to deplete NSD3 and test the effect this has on epithelial cell identity in Read More …
In my last blog post, I observed morphological changes that were indicative of an epithelial to mesenchymal transition (EMT) when NSD3short was overexpressed from a stably integrated transgene in the cancer cell line H1299. Importantly, NSD3 has been identified as amplified in a number of cancer types, including lung (Figure 1) (cBioPortal: Cerami et al., Cancer Discov. 2012 Read More …
Recently, a study of cancer-focused protein-protein interactions identified a direct interaction between NSD3 and the oncoprotein Myc in the non-small cell lung cancer cell line H1299 (Li, Z et al. 2017). Importantly, the study found that increased NSD3 expression levels can activate Myc in H1299 cells as measured by an E-box luciferase reporter. However, the functional significance of Read More …
The early stages of a project often involve spending considerable time building tools to effectively tackle the problem. In cell biology, this often includes testing antibodies and expression constructs in your cell lines of interest. Putting in the work to properly validate these reagents early on in a study is critical. Poorly validated antibodies are, at least Read More …
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 …
