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. Overall design: We have studied poly(A) profiles genome wide using the 3' READS technique in yeast strains harboring precise ORF deletions.

可变聚腺苷酸化（Alternative polyadenylation）可产生一系列具有不同3'端的mRNA同工型；单个基因可生成多种3'端mRNA同工型，并具备典型的多聚腺苷酸（poly(A)）位点使用模式。为鉴定聚腺苷酸化位点分布的潜在决定因素，我们借助CRISPR技术构建了酿酒酵母（Saccharomyces cerevisiae）菌株，该菌株在编码参与基因表达过程中各类因子的候选基因中携带精确的开放阅读框（ORF）缺失。我们采用3' READS技术，对上述突变菌株的全基因组范围内的3' mRNA同工型端点模式进行了测定。整体实验设计：我们利用3' READS技术，对携带精确开放阅读框（ORF）缺失的酵母菌株的全基因组聚腺苷酸化谱进行了分析。