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Analysis of p(A)+ and Spliced RNA in SV40 and Related Constructs

This study investigates the relative amounts of unspliced and spliced p(A)+ RNA in various SV40 constructs, focusing on their exon composition and the impact of specific sequence modifications. The results reveal differences in RNA processing and stability, providing insights into the functional dynamics of RNA in viral contexts. By examining multiple genetic variants, including those incorporating specific mutations, we aim to elucidate the mechanisms governing RNA maturation and expression levels in the SV40 system.

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Analysis of p(A)+ and Spliced RNA in SV40 and Related Constructs

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  1. A D4 A pA SV40 SV-SD4/RRE/SA-pA RRE SA5 BPS 1 A4c BPS 2 A4a A4b PPy PPy * * * * * AAAAAGTGTTGCTTTCATTGCCAAGTTTGTTTCATGACAAAAGCCTTAGGCATCTCCTATGGCAG SA5 AAAAAGTGTTGCTTTCATTGCCACGTTTGTTTCATGACAAACGCCTTCGGCATCTCCTATGGCAG SA5 AG- AAAAAGTGTGGCGTCCGTTGCCACGTTTGTTTACTAACAAACGCCTTCGGCATCTCCTATGGCAG SA5 b1– b2+AG– AAAAAGTGTGGCGTCCGTTGCCACGTTTGTTTACTAACAAACGCCTTCGGCATCTCCTATTTCAG SA5 b1– b2+AG– Py+ AAAAAGTGTGGCGTCCGTTGCCACGTTTGTTTACTAACAAACGCCCTCGCCTTCTCCTCTTTCAG SA5 b1– b2+AG– Py++ AAAAAGTGTGGCGTCCGTTGCCACGTTTGTTTCATGACAAACGCCCTCGCCTTCTCCTCTTTCAG SA5 b1– AG– Py++ AAAAAGTGTTGCTTTCATTGCCACGTTTGTTTCATGACAAACGCCCTCGCCTTCTCCTCTTTCAG SA5 AG– Py++ AAAAAGTGTTGCTTTCATTGCCAAGTTTGTTTCATGACAAAAGCCCTAGCCTTCTCCTCTTTCAG SA5 Py++ B - Rev + Rev SA5 b1-b2+AG-Py++ SA5 b1-b2+AG-Py++ SA5 b1-b2+AG-Py+ SA5 b1-b2+AG-Py+ SA5 b1-AG-Py++ SA5 b1-AG-Py++ SA5 b1-b2+AG- SA5 b1-b2+AG- SA5 AG-Py++ SA5 AG-Py++ SA5 AG- SA5 Py++ SA5 Py++ SA5 AG- SA5 SA5 us s hGH 1 7' 8' 2 3 4 5 6 7 8 1' 2' 3' 4' 5' 6' C 100 100 80 80 60 60 rel. amount of unspliced p(A)+RNA [%] rel. amount of spliced p(A)+RNA [%] 40 40 20 20 0 0 1' 2' 3' 4' 5' 6' 7' 8' 1 2 3 4 5 6 7 8 Exon 1 Exon 2

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