2024-01-17

Presentation by Claire Harbison

Dead Cas13 (dCas13) has been shown to modulate alternative splicing.

A classic and useful example of exon skipping is the SMN2 gene. Two different versions of the SMN2 gene were made with fluoroscent protein on the ends to allow for easier quantification. - DualIN minigene. Inclusion of exon 7 causes dual fluorescence. Exclusion of exon 7 causes only GFP fluorescence. - DualEX minigene. Inclusion of exon 7 causes only GFP fluorescence. Exclusion of exon 7 causes dual fluorescence. gRNAs tiled along the gene to see which guides cause exon 7 to be included or excluded.

Testing. Take dCas13 \(/rightarrow\) DualIN / DualEX \(/rightarrow\) inclusion guide / exclusion guide

1. dCas13 + DualIN + exclusion guide \(/rightarrow\) GFP signal
2. dCas13 + DualIN + inclusion guide \(/rightarrow\) dual signal
3. dCas13 + DualEX + exclusion guide \(/rightarrow\) dual signal
4. dCas13 + DualEX + inclusion guide \(/rightarrow\) GFP signal