Identification of factors influencing the pattern of polyadenylation site use in Saccharomyces cerevisiae

Alternative polyadenylation gives rise to a wide variety of mRNA isoforms with distinct 3' ends; an individual gene can yield many 3' mRNA isoforms and has a typical pattern of poly(A) site use. To identify possible determinants of polyadenylation site distribution, we have used CRISPR to create Saccharomyces cerevisiae strains harboring precise ORF deletions in candidate genes encoding various factors involved in gene expression. We have performed 3' READS to determine the genome-wide pattern of 3' mRNA isoform endpoints in these mutant strains. We have studied poly(A) profiles genome wide using the 3' READS technique in yeast strains harboring precise ORF deletions.