ＣＲＩＳＰＲ／Ｃａｓ９封入高分子集合体の構築と生体内デリバリーへの挑戦

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Cas9 activities of various cationic polymers were evaluated by luciferase-based Cas9 activity system. This assay consists of three different plasmids - sgRNA-expressing plasmid, Stop codon-contained luciferase-expressing plasmid, and luciferase donor DNA plasmid, which were kindly donated by Professor Moritoshi Sato (UTokyo). A cationic polymer (PAsp(OCT/DET)) was complexed with Cas9 messenger RNA and then the resulting polyplexes were transfected into human cervical cancer (HeLa) cells. The Cas9 activities were measured by a luminescence microplate reader. The cationic polymer elicited the similar luminescence intensity compared to commercially available transfection agent, lipofectamine 3000. This result indicates that the new cationic polymer is promising for in vitro Cas9 mRNA transfection.