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Testing RRL-NSD3-Short-3xFLAG-IRES-Puro for Stable Expression in H1299 Cells

descriptionPublicationkeyboard_double_arrow_right Report 09 Feb 2018Publisher:Zenodo

Authors: Dilworth, David; Barsyte-Lovejoy, Dalia;

doi: 10.5281/zenodo.1170481 , 10.5281/zenodo.1170482

Testing RRL-NSD3-Short-3xFLAG-IRES-Puro for Stable Expression in H1299 Cells

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Abstract

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.

SGC Open Notebook Project to Characterize the HMTase NSD3 Exp014 Overview: In this experiment, I tested a lentiviral construct for NSD3-Short-3xFLAG expression. This vector contains an EF1a promoter driving NSD3-3xFLAG-IRES-Puro_Resistance bicistronic transcript (RRL). Also included is an empty vector control and eGFP expression in the same backbone, however lacking the ires-puro resistance sequence. I was unable to select for puromycin resistant cells with the NSD3 expression construct.

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