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Fig. 1 | BMC Genomics

Fig. 1

From: DECKO: Single-oligo, dual-CRISPR deletion of genomic elements including long non-coding RNAs

Fig. 1

Outline of DECKO. a Deletion of genomic elements such as promoters by dual CRISPR targeting. b Structure of pDECKO. Two independent gRNA transgenes are driven by human U6 and human H1 promoter sequences. Variable sequence are highlighted in blue. c Cloning step 1: Synthesised oligonucleotide Insert-1, carrying all variable sequence, is inserted into BsmBI-digested parental plasmid by Gibson assembly. d Cloning step 2: the preassembled universal Insert-2 is inserted into the intermediate plasmid by standard ligation. e Nucleotide sequence of the 165 nt Insert-1. From left to right: Light orange: 5’ Gibson assembly overhang; light blue: targeting region of gRNA1; grey: fragment of first scaffold region; yellow: BsmBI sites; dark blue: target region of gRNA2; dark orange: fragment of H1 promoter; grey: 3’ Gibson assembly overhang

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