 | Figure 1
Nature Genetics
34, 263 - 264 (2003)
Published online: 8 June 2003; | doi:10.1038/ng1173
Induction of an interferon response by RNAi vectors in mammalian cellsAlan J Bridge, Stephanie Pebernard, Annick Ducraux, Anne-Laure Nicoulaz
& Richard Iggo | | | | Figure 1. OAS1 induction by shRNA vectors.
(a) Schematic diagram showing the structure of the shRNA lentiviral vector. (b−f) Quantitative PCR assays. (b−d) Lentiviral infection of HLFs. (b) OAS1 is induced by the vector that targets MORF4L1 (pAB319). (c) Induction of OAS1 does not correlate with MORF4L1 silencing (shRNA 1 is pAB319; the sequence of all five shRNAs is given in the Supplementary Note online). (d) OAS1 induction by MORF4L1 shRNA is not suppressed by restoration of MORF4L1 expression with an uncleavable MORF4L1 cDNA. shRNA: pAB319; cDNA: pAB349 (the cleavage site contains 6 mismatches). (e) Transfection of HLFs with the MORF4L1 siRNA encoded by pAB319 does not induce OAS1. (f) Transfection of HeLa cells with plasmid vectors induces OAS1. The MORF4L1 and MORF4L2 plasmids encode the shRNAs expressed by pAB319 and pAB322. The reduction in MORF4L1 mRNA with the MORF4L2 plasmid is expected from the sequence. (g) Northern blotting for shRNAs targeting MORF4L1 and MORF4L2 of RNA extracted from HLFs infected with lentiviral vectors shows the presence of only the correctly processed 21-nt siRNAs (mock, no virus; blank, pAB303; MORF4L1, pAB319; MORF4L2, pAB322).
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