An in vitro method to induce a precise excision of nucleic acid sequence in a Gene in a cellComprising: (a) selecting a First and a Second Target Double stranded Nucleic Acid In The Genetic Sequence, Understanding each Target Nucleic Acid in a ChainA Ground adjacent to the protoespaciador (PAM) in a 3 extremity (b) Design technically two routing crispr RNA (arncr)Each comprising: - a sequence complementary to a part of the opposite Chain of the target Nucleic Acid that doesnt understand the reason, PamAnd - a sequence of 3 extension (c) providing at least one routing crispr RNA transactivator (arncrtra) comprising a sequence complementary to a part of the sequences3 extension of such arncr b) (d) providing at least one nickasa cas9 is a Nuclease that houses a Nuclease Domain cas9 ruvc type non Functional or non functional and that recognizes HNHThe Reason (or reasons) Pam (e) introducing into the cell (such arncr, such or such and such nickasa) arncrtra cas9 where the cell is a T Lymphocyte.So that each Complex cas9 - arncrtra: arncr induces a notch in the event of double stranded Nucleic Acid targets to Cleave the DNA Sequence between the first and second \u00e1ci targetsOJ nucleic.Un método in vitro para inducir con precisión una escisión de ácido nucleico en una secuencia genética en una célula, que comprende: (a) Seleccionar una primera y una segunda diana de ácido nucleico bicatenario en dicha secuencia genética, comprendiendo cada diana de ácido nucleico, en una cadena, un motivo adyacente al protoespaciador (PAM) en una extremidad 3 (b) diseñar técnicamente dos ARN de direccionamiento de CRISPR (los ARNcr), comprendiendo cada uno: - una secuencia complementaria a una parte de la cadena opuesta de la diana de ácido nucleico que no comprende el motivo PAM, y - una secuencia de extensión 3 (c) proporcionar al menos un ARN de direccionamiento de CRISPR transactivador (ARNcrtra) que comprende una secuencia complementaria a una parte de las secuencias de extensión
