Targeting N-Terminal Huntingtin with a Dual-sgRNA Strategy by CRISPR/Cas9

Figure 3

Targeting N-terminal HTT with a dual-sgRNA strategy. (a) Design of sgRNAs flanking the polyQ region. (b) Detection of cleavage efficiencies with different combinations of sgRNA pairs by the T7E1 assay. Mixed indels were calculated based on the fractions of PCR products. “a” represents the undigested PCR product, and “b” and “c” represent cleavage products. (c) Single colonies screening by junction PCR after sgHTT-4/9 targeting. (d) Sanger sequencing results of gDNA and cDNA junctions in the sgHTT-4/9-#7 cell colony. (e) Measurement of HTT levels by quantitative PCR. (f) Western blot detection of full-length HTT and possible truncated HTT proteins.

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