Compositions and methods for binding to a target sequence of interest are provided. The compositions find use in cleaving or modifying a target sequence of interest, visualization of a target sequence of interest, and modifying the expression of a sequence of interest. Compositions comprise RNA-guided nuclease (RGN) polypeptides, CRISPR RNAs, trans-activating CRISPR RNAs, guide RNAs, and nucleic acid molecules encoding the same. Vectors and host cells comprising the nucleic acid molecules are also provided. Further provided are RGN systems for binding a target sequence of interest, wherein the RGN system comprises an RNA-guided nuclease polypeptide and one or more guide RNAs.

Novel engineered non-natural deaminases are provided. Methods and compositions are provided for modifying the genome of a cell, including a plant cell. Genome modification through guided CRISPR polypeptides and multiple deaminases are provided. Engineered non-natural novel deaminases, including adenosine deaminases are provided. These novel deaminases have improved activity in cells including plant cells.

The present disclosure relates to Neisseria meningitidis (Nme) 2 Cas9 (Nme2Cas9) and Nme2.sup.SmuCas9 variants comprising one or more amino acid substitutions with increased genome editing activities (e.g., improve nuclease and base editing efficiencies).

Provided herein are fusion polypeptides comprising a Cpf1 domain lacking nuclease activity and an endonuclease domain. Also provided herein are fusion polypeptides further comprising a genomic modification domain, which in some embodiments is a base editor, such as a deaminase. Also provided herein are methods involving contacting the fusion polypeptides with a gRNA to form a genetic editing system directed to a target site sequence in the genome of a cell.

The present disclosure relates to a composition for gene editing and a use thereof. According to the present disclosure, an adenine base editor LbABE8e system in which a Cas12a protein variant including one or more mutations and adenine deaminase are fused has altered PAM specificity to achieve a significant genomic single base editing effect on target DNA, and thus expands the range of applicable CRISPR system selection. Accordingly, due to gene editing, it is expected that the adenine base editor LbABE8e system may be widely used in various fields, such as gene therapy, creation of commercial profits with industrial strains with improved productivity, improvement of the quality of public health care through improvement of intestinal microorganisms, and improvement of crops and livestock breeds free from GMO issues.

When a cancer patient is administered an anti-cancer therapy targeting a lineage specific cell-surface antigen (e.g., CD33 (Siglec-3), CLL-1, CD123, CD327 (Siglec-6), and/or CD312 (EMR2)), e.g., in the form of an immunotherapeutic agent, the therapy can deplete not only cancer cells expressing the lineage-specific cell-surface antigen, but also noncancerous cells expressing the lineage-specific cell-surface antigen in an on-target, off tumor effect. This disclosure provides, e.g., novel cells having a modification (e.g., insertion or deletion) in an endogenous lineage-specific cell-surface antigen (e.g., CD33 (Siglec-3), CLL-1, CD123, CD327 (Siglec-6), and/or CD312 (EMR2)) gene. The disclosure also provides compositions, e.g., gRNAs, that can be used to make such a modification.

The present disclosure relates to Cast 2a effector proteins with increased activity compared to previously described Cas12a effector proteins. The present disclosure also relates to fusion proteins comprising a Cas12a effector protein fused to a deaminase with increased activity compared to previously described fusion proteins comprising a Cast 12a effector protein fused to a deaminase. Systems and methods of their use are also disclosed.

The present invention provides improved methods for generating banana embryos and plants carrying desired mutations in target sequences, comprising introducing at least one mutation in at least one endogenous acetolactate synthase (ALS) gene. The invention also provides banana plants and products generated by such methods.

Compositions and methods comprising novel deaminase polypeptides for targeted editing of nucleic acids are provided. Compositions comprise deaminase polypeptides. Also provided are fusion proteins comprising a DNA-binding polypeptide and a deaminase of the invention. The fusion proteins include RNA-guided nucleases fused to deaminases, optionally in complex with guide RNAs. Compositions also include nucleic acid molecules encoding the deaminases or the fusion proteins. Vectors and host cells comprising the nucleic acid molecules encoding the deaminases or the fusion proteins are also provided.

An immune cell that is genetically engineered to express an exogenous alternative carbon source (ACS) metabolism gene, in which the ACS is not glucose and wherein the ability of the immune cell to metabolise the ACS is increased due to expression of the exogenous ACS metabolism gene. Also provided are polynucleotides, vectors, pharmaceutical compositions, methods of genetically engineering the immune cell and methods of use in therapy.