Abstract: The disclosure provides AAV expression cassettes for production of AAV viral vectors, wherein the expression cassettes comprise a first inverted terminal repeat (ITR); a first promoter; a first gRNA comprising a first gRNA targeting region; a second promoter; a second gRNA comprising a second gRNA targeting region; a third promoter; a third gRNA comprising a third gRNA targeting region; and a second ITR. The disclosure also provides AAV viral vectors, including self-complimentary AAVs, comprising the expression cassettes of the disclosure. The AAVs disclosed herein may be used to treat genetic diseases, such as Duchenne Muscular Dystrophy (DMD).