Skip to main content
. 2014 Nov 20;43(3):e17. doi: 10.1093/nar/gku1198

Figure 1.

Figure 1.

Knockout of miR-21 in HEK293 cells. (A) Strategy for making Cas9 all-in-one vector targeting miR-21 precursor. Three gRNA target regions are shown at the top panel; a single miR-21 gRNA each was cloned into the vector at the bottom panel as detailed in the Materials and Methods section. (B) Detection of indels in miR-21 knockout cells by T7 endonuclease 1 or Surveyor nuclease assay, as detailed in the Materials and Methods section. The black arrow indicates wild-type band and the red arrows indicate mutant bands, with relative cleavage frequency shown at the bottom. (C) A donor vector for targeting miR-21 through homologous recombination (HR). pA, poly A signal. (D) A procedure of selection and identification of knockout clones. (E) miR-21 knockout increased colony number, as determined by colony formation assay for a mixed pool after puromycin selection. Values are means of ± SE (n = 3). **P < 0.01.