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. 2023 Jun 17;24(12):10258. doi: 10.3390/ijms241210258

Figure 4.

Figure 4

Spliced mRNA before and after rAI insertion and treatment with recombinase. Idealized mRNA after splicing, (A) before any insertion, and (B) after rAI insertion, followed by recombinase treatment so that the rAI no longer has its branchpoint and is unlikely to splice itself out. It is best to choose a constitutive exon in which one can follow the rules for rAI placement within an exon and one that is close to but not within 200 nt downstream of the start codon and is upstream from another constitutive exon. This preferred location is shown with the dark blue triangle. The closer to but not within 200 bp of the start codon, the greater the chance of mRNA-degradative processes occurring after recombinase treatment. In this example, an rAI non-divisible by 3, such as SCON, is demonstrated, causing a frameshift leading to multiple downstream premature termination codons (PTCs). This allows for a greater chance of mRNA degradation through NMD and by creating a long 3′ untranslated region (long 3′ UTR).