Figure 1.

Schematic drawing of the Cas9 expression cassette and the RNA polymerase III U6 promoter (light green; P_U6) driven sgDNA (pSM2) according to Schuster et al., 2016 [23]. Yellow: 19 bp target sequence; dark green: scaffold sequence; black: U6 terminator; light purple: constitutive otef-promoter (P_otef); light gray: nuclear localization signal (NLS); purple: open reading frame (ORF) of high fidelity cas9; dark gray: HA-tag; dark purple: nos terminator. The former XbaI-site inside of the U6 terminator was inactivated by a point mutation. The 32/36 bp flanking sequences for the oligonucleotides are given.