Fig 5. Production of truncated MALAT1 RNA from mutated alleles.

(A) RT-PCR was performed on RNA from bulk cells where MALAT1 exon region was deleted (sgRNA Pair 1, in two biological replicates), or control cells transfected with pDECKO targeting EGFP. Primers flanking the deleted region were used, and are expected to amplify fragments of the indicated sizes, depending on whether the RNA arises from a wild type or a deleted allele. Specificity was ensured by the exclusion of the reverse transcriptase enzyme in control reactions (“RT-”). (B) Sequencing analysis of mutant junctions of 4 of the colonies after TA cloning of the RT-PCR product. In red, region complementary to the sgRNA variable region; Green, PAM sequences; Blue, indel. Expected cut location is marked with vertical bar.