Abstract: The present disclosure relates to compositions and methods for simultaneous and combinational CRISPR-mediated gene activation, gene editing, and gene repression in plants based on a single Cas protein.

Abstract: Compositions and methods for modifying genomic DNA sequences of a plant cell are provided. The methods produce double stranded breaks at target sites in a genomic DNA sequence, resulting in mutation, insertion, and/or deletion of DNA sequences at the target site(s) in a genome. The compositions comprise DNA constructs comprising nucleotide sequences that encode a Cas12b protein. The DNA constructs can be used to direct the modification of genomic DNA at a target site. Methods to use these DNA constructs to modify genomic DNA sequences are described herein. Additionally, compositions and methods for modulating the expression of genes are provided.

Abstract: A non-naturally occurring heterologous CRISPR-Cas12a genomic editing system is described, including or encoding at least one Cas12a ortholog endonuclease selected from among Lb5Cas12a, CMaCas12a, BsCas12a, BoCas12a, M1Cas12a, Mb2Cas12a, MbCas12a TsCas12a, and MAD7 endonucleases. The ortholog endonucleases enable genomic editing of plants at low temperatures, e.g., below 25° C. The CRISPR-Cas12a system can be employed for targeting PAM sites such as TTN, TTV, TTTV, NTTV, TATV, TATG, TATA, YTTN, GTTA, and GTTC, utilizing corresponding gRNAs, and is readily adapted to multiplexed editing operations of plants such as rice and other food crops.