SGC Open Notebook Project to Characterize the HMTase NSD3 Exp016 Objective: Bicistronic elements are common features in lentiviral expression systems that allow expression of two protein products from a single promoter. The two most common bicistronic elements are the internal ribosome entry site (IRES), which allows cap-independent translation of a second open reading frame within an mRNA, and the 2A system, which introduces a self-cleaving peptide between two desired protein products. There are several advantages to the the 2A system over an IRES, which include matched expression levels of your two protein products of interest as well as having a smaller footprint (~1/10 size in kb), which may improve viral titre. Therefore, I will use site-directed mutagenesis to alter the NSD3short lentiviral expression plasmid to replace the IRES sequence with a T2A sequence.

Funding Acknowledgment: The SGC is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada through Ontario Genomics Institute [OGI-055], Innovative Medicines Initiative (EU/EFPIA) [ULTRA-DD grant no. 115766], Janssen, Merck KGaA, Darmstadt, Germany, MSD, Novartis Pharma AG, Ontario Ministry of Research, Innovation and Science (MRIS), Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and Wellcome.