Figure 3.

CRISPR-Cas9 Editing Reveals that miR-10b Expression Is Essential for Glioma Viability

(A) Light microscopy images of glioma cells transfected with either the control empty vector or the miR-10b-targeting vectors demonstrate the appearance of floating apoptotic cells in the edited cultures (upper panels). Schematic view of the analysis of miR-10b DNA locus in the floating cells. The DNA was isolated from the sgRNA nG1/G3-targeted cultures, and the miR-10b genomic locus was amplified and sequenced. The sequencing results reveal a range of miR-10b mutants, with 17 of 20 clones mutated in the miR-10b locus. (B) Surveyor cleavage assay of the attached and floating populations of LN229 and U251 glioma cells demonstrates that miR-10b is edited preferentially in floating cells, whereas the unedited cells remain attached. (C) miR-10b levels are reduced in the floating apoptotic, but not in the attached viable, LN229 cells.