Figure 1 | Scientific Reports

Figure 1

From: Efficient genetic manipulation of the NOD-Rag1-/-IL2RgammaC-null mouse by combining in vitro fertilization and CRISPR/Cas9 technology

Figure 1

gRNAs targeting Fah exon5 can efficiently direct Cas9 cleavage of target DNA.

(a) gRNA sequences against Fah exon5. The exon 3, 4, 5 and 6 region of the mouse Fah gene is shown. The exon5 sequence (upper case) and part of the upstream intron sequence (lower case) are shown with 3 gRNA sequences (labeled in red) and the PAM sequence NGGs in turquoise. (b) and (c) gRNA cleavage efficiency in vitro. (b) Schematic diagram of gRNA mediated Cas9 DNA cleavage. The PCR DNA product containing the gRNA target sites, Cas9 nuclease protein and gRNA are mixed. The Cas9/gRNA complex cuts the PCR product into two fragments. (c) The cleaved DNA was resolved on TapeStation. A0(L), DNA ladder; A1, PCR DNA alone; B1, PCR DNA + Cas9 nuclease; C1, D1 and E1, PCR DNA + Cas9 nuclease + gRNA1 (C1), gRNA2 (D1) and gRNA 3 (E1). Full-length gel is shown. The gRNA mediated cutting efficiency is shown below each lane.

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