Nanopore Sequencing Analysis of Early Erythroid Cells Following PABPC1 Knockdown
Erythropoiesis is precisely regulated by a multi-layered network, crucial for maintaining stable hemoglobin levels and ensuring effective oxygen delivery. Selective polyadenylation (APA) is a post-transcriptional regulatory mechanism, generating various messenger RNA subtypes from a single gene based on specific 3'-untranslated region sequences. Although APA plays a vital role in various cellular processes, its mechanisms in erythropoiesis remain to be explored. In this study, we initially found that knocking down the key APA regulatory protein PABPC1 disrupts the proliferation and differentiation of erythroid progenitor cells. To further elucidate its mechanism, we performed nanopore sequencing (ONT sequencing) on early erythroid cells treated with PABPC1 lentiviral knockdown. Overall design: We collected three samples of human mobilized peripheral blood, isolated CD34+ cells using magnetic beads, and induced them to differentiate into red blood cells in vitro. At the early stage of erythroid differentiation, we performed lentiviral knockdown treatments with shNC/shPABPC1 (shPABPC1_1_1, shPABPC1_1_2, shPABPC1_1_3 vs shNC_1, shNC_2, shNC_3), and 48 hours later, we collected the treated cells and extracted RNA for sequencing.