Figure 2

Insertion of alternative termination signals fail to reduce the transcriptional readthrough activity of LV SIN LTR. A. Schematic diagram of lentiviral SIN LTRs containing various termination signal elements. HTLVpA (a 130 bp fragment from MT4 cell genomic DNA) and hGH intron (a 193 bp fragment from pXO-hGH) were PCR amplified and cloned into pdl-EF-3'LTR-IRES-nlCre. The mutated tRNA (mu-tRNA, 236 nt) sequence was generated by annealing several synthetic oligos and cloned into LV SIN vector. All amplified fragments were verified by sequencing of the subclones and the final reporter clones. B. Quantitative analysis of termination signal insertion on SIN LTR readthrough. One-way ANOVA analysis reveals a significant difference between WT and the other groups (p < 0.05). No significant difference is detected between the different chimeric termination signal SIN LTR constructs. The modest difference seen between the chimeric termination signal SIN LTRs and the SIN LTR is significant (p < 0.05, marked by asterisk).