Using pgRNA–Cas9 system to knockout MKL1 inhibited cell migration and proliferation
2018
Liu, Hong-Bo | Yin, Ze | Zheng, Peng | Xu, Yao | Wang, Zhen-Yu | Li, Xi | Zhang, Tong-Cun | Lim, Junxian
As a transcription factor, megakaryoblastic leukemia 1 (MKL1) binds with serum response factor (SRF) to regulate targeted genes expression. Therefore, it involved in various cellular processes, such as cancer cells migration, proliferation and differentiation. However, the report about knockout MKL1 expression using CRISPR–Cas9 system in human genome is rare. Therefore, paired-guide RNA (pgRNA) library was constructed, and pgRNA–Cas9 system was used to delete MKL1 expression in HeLa cells. The result showed that the expression of MKL1 was decreased 95% by Western blotting. And wound healing assay and MTT assay indicated that depletion MKL1 inhibited cell migration and proliferation. Additionally, the protein expression levels of tumor suppressor factors p21, p53, pRB and DLC1 were changed after depletion MKL1. These data suggested that tumor suppressor factors p21, p53, pRB and DLC1 maybe played an important role in the cell growth and migration of MKL1-depletion cells.
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