Jaak2 Biogenesis - 1866 Words

JAK2/STAT signaling pathway was one of the most significantly differentially expressed gene sets in BC compared with CP progenitors. This signaling pathway increases the ADAR1 expression in progenitor LSCs and promotes ADAR1-mediated A-to-I editing. Furthermore, ADAR1 impairs biogenesis of tumour suppressive miRNAs, thereby contributing to cancer progression. Since the let-7 miRNA family is also related to self renewal of LSC, it led the authors to investigate the correlation between JAK2/STAT pathway and LIN28B/let-7-axis, they hypothesized that the LIN28B/let-7self-renewal axis may also be disrupted by increased JAK2 signaling. This hypothesis was validated by a lentiviral transduction of BCR-ABL1 in normal progenitor, it shows an†¦show more content†¦An increase in LIN28B expression can promote cancer progression due to the inhibition of let-7 miRNAs. 6. In the experiment where the authors quantify the JAK2 expression after transducing CD34+ progenitor cells (figure 2B-D), please identify all the controls and normalizations used and briefly describe their significance. Use  ¾ of a page maximum. (5.5 marks) Control The control to quantify the JAK2 expression after transducing CD34+ progenitor cells is the lentiviral vector backbone, which is involved in the lentiviral JAK2 transduction into the human JAK2-GFP in normal CD34+ progenitors. Fluorescence measurement is compared between CD34+ cord blood cells transduced with lentiviral vector backbone and the normal CD34+ cells, in order to examine the transduction efficiency. JAK2 expression of unaffected genes without the transgene is observed, acting as a comparison to the overexpression JAK2 in the transduced CD34+ progenitor cells (1). Normalization Lentiviral ADAR1 transduction efficiency of LSC is measured by luciferase reporter activity, which is plotted against pCDH lentiviral vector and pDEST26 mammalian expression vector. Transduction efficiency is measured and normalized by fluorescence intensity of p-JAK2 and p-STAT5a, and JAK2 qRT-PCR. In the qRT-PCR analysis, a housekeeping gene RPL27 (ribosomal protein