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Designing a new artificial transcription factor based on engrailed homeodomain

Sunami, Tomoko; Kono, Hidetoshi

Recently, zinc finger nucleases which are artificially fused a zincfinger transcription factor and a nuclease have been widely used for genome editing. However, the applications are limited because they only recognize GC-rich sequences. To make such artificial proteins recognize broader sequences other than GC-rich ones, we are rationally designing novel proteins based on an engrailed homeodomain. Engrailed homeodomain proteins recognize six bases. To recognize longer DNA sequences, we connected two homeodomains with a short linker. We will report that the designed molecule show a strong affinity and high specificity for the target sequence. We will also report a DNA sequence profile of the protein obtained by B1H assay.

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