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    • 8. 发明申请
    • METHODS TO REPROGRAM SOMATIC CELLS TO ALTERNATIVE CELL FATES OR PRIMITIVE CELL STATES
    • US20220356451A1
    • 2022-11-10
    • US17597776
    • 2020-07-23
    • Tania RayPartha S. Ray
    • Tania RayPartha S. Ray
    • C12N5/077C12N15/113
    • Induced overexpression of defined exogenous transcription factors (TFs), or alternatively treatment with specific pathway modulatory cocktails, can reprogram somatic cells to pluripotency or alternate cell states. A barrier to initiating reprogramming lies in the starting cell's molecular identity, enforced by lineage-instructive TFs. However, it remained unclear whether repression of such somatic lineage-defining TFs of the starting cell in the absence of exogenous TFs is sufficient to induce cell reprogramming Using an intra-species somatic cell hybrid model, SNAI2 and PRRX1 were identified as the most critical determinants of mesenchymal commitment in rat embryonic fibroblasts (REFs) and demonstrate that siRNA-mediated transient knockdown of these individual factors is adequate to convert REFs into functional adipocytes, chondrocytes or osteocytes without requiring the provision of exogenous TFs. Additionally, it was shown that siRNA-mediated transient knockdown of SNAI2 alone, in the absence of exogenous TFs, is sufficient to transform REFs to a dedifferentiated pluripotent stem-like cell (dPSC) state that forms embryoid bodies and is capable of triple germ layer differentiation. These results establish for the first time that transient repression of a single somatic lineage-defining TF can effectively induce transdifferentiation to alternative somatic cell states or dedifferentiation to dPSCs in the absence of exogenous TFs or small molecule cocktails.