A20-eGFP-Neo/Fluc-Puro

Species: Mouse (Mus musculus)

Cell type: Lymphoma

Reporter genes: Enhanced green fluorescent protein (eGFP), Firefly luciferase (Fluc)

Media: RPMI, 50 mM b-mercaptoethanol, 10% fetal bovine serum (FBS), 1% Penicillin/Streptomycin

Description: 

This is a cell line derived from the murine lymphoma A20 cell line (ATCC® TIB-208^TM). Parental A20 cells were transduced with 1) LV-eGFP-P2A-Neo (Imanis #LV067) encoding the enhanced green fluorescent (eGFP) cDNA under the spleen focus-forming virus (SFFV) promoter and linked to the neomycin resistance gene (Neo) via a P2A cleavage peptide and 2) LV-Fluc-P2A-Puro (Imanis #LV012) encoding the firefly luciferase (Fluc) cDNA under the SFFV promoter and linked to the puromycin resistance gene (Puro) via a P2A cleavage peptide. A high Fluc and eGFP expressing population was generated by selection using puromycin and G418 followed by selection using a methylcellulose based semi-solid medium. The lentiviral vectors are self-inactivating (SIN) vectors in which the viral enhancer and promoter have been deleted. Transcription inactivation of the LTR in the SIN provirus increases biosafety by preventing mobilization by replication competent viruses and enables regulated expression of the genes from the internal promoters without cis-acting effects of the LTR¹.

¹Miyoshi et al. J Virol. 1998. 72:8150-8157.