cDNA sequencing of TP53-mRNA transcripts

The aim of this part of the study was to determine whether wild-type p53 and its missense mutant variant R248Q have comparable or different quantities of mRNA within the cell (using isogenic MOLM13 AML cell lines modified by CRISPR-Cas9 engineering to observe TP53 allelic series), which would resolve whether R248Q accumulation within cancer cells (such as acute myeloid leukemia cells) is caused by an increased transcription rate from the missense mutant TP53 allele compared to the wild-type TP53 allele. Therefore, we observed and compared MOLM13 cells with one TP53 wild-type allele and a TP53 null mutation on the other allele (+/-), MOLM13 cells carrying one R248Q missense mutated allele while having a TP53 null mutation on the other allele, and MOLM13 cells with one wild-type TP53 allele and one R248Q missense mutated allele.