RNA guide genome editing in citrus using CRISPR-ribonucleoprotein complexes

A method of altering expression of at least one gene product comprising introducing into a citrus plant cell an engineered, non-naturally occurring gene editing system comprising one or more vectors, said citrus plant cell containing and expressing a DNA molecule having a target sequence present within a gene encoding the at least one gene product, said gene editing system comprising:

• (a) a first regulatory element operable in a plant cell operably linked to at least one nucleotide sequence encoding a CRISPR-Cas system guide RNA (gRNA) that hybridizes with the target sequence, and (b) a second regulatory element operable in a plant cell operably linked to a nucleotide sequence encoding a Type-II CRISPR-associated nuclease, wherein components (a) and (b) are located on same or different vectors of the system, whereby the guide RNA targets the target sequence and the CRISPR-associated nuclease cleaves the DNA molecule, whereby expression of the at least one gene product is altered;
• and, wherein the CRISPR-associated nuclease and the guide RNA do not naturally occur together;
• wherein said first regulatory element comprises SEQ ID NO:
• 6 or a nucleotide sequence having at least 98% nucleotide sequence identity to the nucleotide sequence as set forth in SEQ ID NO:
• 6;
• and wherein said class II CRISPR-associated nuclease comprises Cas9 or cpf1.