96 The Use of a Mutant RNA Polymerase II Largest Subunit That Confers α Amanitin Resistance as a Selection Marker for Ex Vivo Gene Therapy Molecular Therapy �������� ��� ���� ���������������� �������[.]
RNA VIRUS VECTORS I 94 Recombinant Sendai Virus-Mediated CFTR cDNA Transfer 95 Early Detection of 2-LTR Junctions in MoMLV Infected Cells Cytoplasm Stefano Ferrari,1,4 Raymond Farley,1,4 Felix Munkonge,1,4 Uta Griesenbach,1,4 Stephen N Smith,1,4 Jun You,2 Tsuyoshi Tokusumi,2 Akihiro Iida,2 Brandon Wainwright,3 Mike Gray,5 Angela Wright,5 Bernard Verdon,5 Barry Argent,5 Duncan M Geddes,1,4 Mamoru Hasegawa,2 Eric W F W Alton.1,4 Gene Therapy, NHLI,Imperial College, London, United Kingdom; 2DNAVEC Research Inc, Tsukuba, Ibaraki, Japan; Department for Molecular & Cellular Biology, University of Queensland, Brisbane, Australia; 4UK Cystic Fibrosis Gene Therapy Consortium, United Kingdom; 5School of Cell & Molecular Biosciences, Newcastle University, Newcastle, United Kingdom Fatima Serhan,1 Magalie Penaud,1 Caroline Petit,2 Ghislaine Duisit,1 Pierre Sonigo,2 Philippe Moullier.1 Inserm ERM 01-05, CHU-Hotel Dieu, Nantes, France; 2Inserm U567, Institut Cochin, Paris, France Recombinant Sendai virus (SeV) can deliver reporter and therapeutic genes to the airway epithelium very efficiently To assess applicability for Cystic Fibrosis (CF), we cloned the human CFTR cDNA into the SeV genome between the Matrix and the Fusion genes CFTR activity was assessed on C127-/- using a radioactive iodide efflux assay and results expressed as change in the apparent rate constant per minute (Δk min-1) ± s.e.m Positive controls (T84 cells) showed maximal efflux rate after forskolin/IBMX (F/I) addition (0.24 ± 0.02, n=30) compared to C127-/- values (0.004 ± 0.004, n=11), p