Method for RGEN RNP delivery using 5′-phosphate removed RNA

A method of decreasing expression of a gene involved in a Type I interferon response following delivery of an RNA-guided endonuclease ribonucleoprotein into a cell wherein the cell comprises the gene involved in the Type I interferon response, the method comprising

• preparing a guide RNA having a 5′ end free of both a diphosphate residue and a triphosphate residue by in vitro transcription using a prokaryote RNA polymerase followed by removing the diphosphate and the triphosphate residues of the guide RNA,
• forming the RNA-guided endonuclease ribonucleoprotein by combining a Cpf1 protein with the guide RNA having the 5′ end free of both the diphosphate residue and the triphosphate residue, administering the RNA-guided endonuclease ribonucleoprotein directly to the cell, and
• wherein the gene involved in the Type I interferon response comprises a gene selected from the group consisting of IFN-β (Interferon-beta), RIG-1 (retinoic acid-inducible gene 1), and OAS2 (2′-5′-oligoadenylate synthetase 2) genes, and the expression level of the gene involved in the type I interferon response is decreased compared to a control in which a RNA-guided endonuclease ribonucleoprotein comprising a Cpf1 protein and a guide RNA having the diphosphate residue or the triphosphate residue at the 5′ end is administered.