Project description:In order to explore the role of LHX2 in neural fate determination, we performed knockdown and overexpression studies in H9. --doxycycline induced LHX2 overexpression in H9-- GFP as a control 3 days doxycyline treatment was performed samples was collected 1 day (Day4) and 13 days (Day16) post doxycyline treatment --shLHX2 in H9-- shLuc as a control samples were collected 12 days of neural induction all samples were biological duplicate
Project description:Mocetinostat (MGCD) which is a kind of histone deacetylase inhibitors (HDACi) promotes human embryonic stem cells (hESCs) differentiation towards neural progenitor cells (NPCs). Application of HDAC inhibitors (HDACi) increased the expression of neuroectodermal markers once neural differentiation was initiated, thereby leading to more NPC generation. We used microarrays to detail the global gene expression during NPC differentiaton upon MGCD treatment and identified the transcript changes effected by MGCD during this process. Undifferentiated H9 hESCs (D0), as well as H9 cells with or without MGCD treatment on day 3 (D3C and D3M) and on day 7(D7C and D7M) of NPC generation were collected for RNA extraction and hybridization on Affymetrix microarrays.
Project description:We have sequenced miRNA libraries from human embryonic, neural and foetal mesenchymal stem cells. We report that the majority of miRNA genes encode mature isomers that vary in size by one or more bases at the 3’ and/or 5’ end of the miRNA. Northern blotting for individual miRNAs showed that the proportions of isomiRs expressed by a single miRNA gene often differ between cell and tissue types. IsomiRs were readily co-immunoprecipitated with Argonaute proteins in vivo and were active in luciferase assays, indicating that they are functional. Bioinformatics analysis predicts substantial differences in targeting between miRNAs with minor 5’ differences and in support of this we report that a 5’ isomiR-9-1 gained the ability to inhibit the expression of DNMT3B and NCAM2 but lost the ability to inhibit CDH1 in vitro. This result was confirmed by the use of isomiR-specific sponges. Our analysis of the miRGator database indicates that a small percentage of human miRNA genes express isomiRs as the dominant transcript in certain cell types and analysis of miRBase shows that 5’ isomiRs have replaced canonical miRNAs many times during evolution. This strongly indicates that isomiRs are of functional importance and have contributed to the evolution of miRNA genes
Project description:We reported loss of ARID1A promoted neurogenesis and inhibited cardiogenesis. Under microscopy, we observed that spontaneously differentiated cells were induced in ARID1A KO H9 hESCs cultured in mTesR medium. After cardiac differentiation for 10 days, we also observed the cell types were totally different between WT and ARID1A KO cells. We did not know what cells types were. Here scRNA-seq were used to identify the cell types in WT H9 hESCs and ARID1A KO H9 hESCs.
