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. 2012 Dec 31;7(12):e52798. doi: 10.1371/journal.pone.0052798

Figure 1. Generation of GLuc.

Figure 1

-expressing patient-derived ALL cells. a Scheme of the lentiviral vector construct; arrow indicates start of transcription; RSV/5′LTR = hybrid of the Rous Sarcoma virus promoter and the U5 long terminal repeat from HIV-1 virus; EF1 P = constitutive elongation factor 1-alpha promoter; GLuc = membrane anchored form of the Gaussia luciferase (GLuc) enzyme fused to the transmembrane domain of CD8; T2A = “self-cleaving” 2A peptide from insect virus Thosea asigna; copGFP = green fluorescent protein cloned from copepod Pontellina plumata; 3′ΔLTR = HIV-1 virus long terminal repeat with a self-inactivating U3 deletion; b, cTransduction efficiency as determined by flow cytometry measurement of GFP expression after one round of amplification of transduced cells in mice; (b) in ALL-50 and ALL-199; (c) in all 9 patient-derived ALL samples studied; d Stability of biological characteristics of patient-derived ALL cells despite of lentiviral transduction; examples from data shown in detail in Supplemental Figure 1; comparison of ALL-199 cells before and after lentiviral transduction and sorting concerning drug-induced cell death after 48 hours in vitro (left panel), expression of cell surface markers (middle panel) and time to engraftment (right panel).