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Fig. 1 | Genome Biology

Fig. 1

From: Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds

Fig. 1

New Cas12a orthologs harnessed to robustly edit mammalian genomes. a Alignment of direct repeats (DRs) from the 25 Cas12a family proteins. Note that 4 Cas12a bacterial loci contain no DR array. Non-conserved bases in the loop are colored, and the conserved stem duplex is highlighted in pink. b (Upper) schematic showing the sequence of crRNA targeting the human CD34 gene site 1. (Lower) T7EI analysis of targeted indel frequencies induced by the 6 Cas12a candidates (ArCas12a, BsCas12a, HkCas12a, LpCas12a, PrCas12a, and PxCas12a) as indicated. AsCas12a was used as positive control. GFP, an empty backbone vector without Cas12a protein expression. M, DNA marker. c (Upper) schematic showing the sequence of crRNA targeting the human CD34 gene site 8. (Lower) T7EI assay result indicated that HkCas12a facilitated targeted indels directed by 5′-TCTN PAM. GFP, an empty backbone vector without Cas12a protein expression. M, DNA marker

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