ENGINEERED CRISPR/CAS9 SYSTEMS FOR SIMULTANEOUS LONG-TERM REGULATION OF MULTIPLE TARGETS

The invention provides CRISPR-based compositions and methods comprising non-repetitive sgRNA promoter and handle sequences for simultaneous, stable expression of multiple sgRNAs. In one embodiment, the invention relates to a nucleic acid molecule comprising an extra long sgRNA array (ELSA) for expression of at least two sgRNA sequences comprising: nucleotide sequences encoding two or more sgRNA sequence, wherein each sgRNA encoding nucleotide sequence is under the control of a sgRNA promoter and operably linked to a sgRNA handle sequence; wherein the ELSA comprises a maximum shared repeat length of 20 nucleotides or less. In one embodiment, the ELSA comprises a maximum shared repeat length 12 nucleotides or less. L'invention concerne des compositions à base de CRISPR et des procédés comprenant un promoteur de ARNsg non répétitif et des séquences pilote pour l'expression simultanée et stable de multiples ARNsg.