The present disclosure provides adenine base editors (ABEs) that have context specificity, i.e., a preference for a pyrimidine positioned 5 of the target adenosine, or preference for a purine positioned 5 of the target adenosine. In addition, methods for targeted nucleic acid editing are provided. Further provided are pharmaceutical compositions comprising the ABEs. Also provided are vectors useful for the generation and delivery of the ABEs, including vector systems for engineering the ABEs through directed evolution. Cells containing such vectors and ABEs are also provided. Further provided are methods of treatment and uses comprising administering the ABEs.

In the various aspects and embodiments, the present disclosure provides cell populations or cell banks thereof to provide immune compatible, allogeneic cell therapies. In the various aspects and embodiments, the cell populations and progeny thereof maintain sufficient HLA Class I and HLA Class II functionalities, while facilitating patient matching to prevent or reduce graft versus host disease (GVHD) or graft rejection. The disclosure further provides methods for creating the populations by gene editing, and methods for cell therapy involving cells or tissues derived from the cell populations (including but not limited to hematopoietic stem cells, or HSCs, progenitors, or progenies thereof).

The present disclosure unravels novel mechanistic information involving the direct regulation of OBSCN via OBSCN-AS1 through chromatin remodeling and enhanced RNA polymerase II recruitment. Remarkably, it is shown herein that targeting of OBSCN-AS1 is sufficient to restore OBSCN expression in highly aggressive triple-negative breast cancer cells. Provided herein are methods for increasing OBSCN expression in a cell, comprising providing to the cell one or more agents that increases levels of OBSCN-AS1 IncRNA or a variant thereof in the cell, and CRISPR/Cas9 systems for increasing levels of OBSCN-AS1 IncRNA in cells.

A protein is provided that consists of a sequence including any one amino acid sequence of the following (a) to (c), forms a homodimer, and forms a complex with a guide RNA: (a) An amino acid sequence containing at least one substitution of an amino acid residue selected from the group consisting of I118, Y122, I126, and M178 in an amino acid sequence set forth in SEQ ID NO: 1, (b) An amino acid sequence in which one to several amino acids are deleted, inserted, substituted, or added in a portion other than amino acid positions 118, 122, 126, and 178 of the amino acid sequence represented by (a) above, (c) An amino acid sequence having 80% or more identity in a portion other than the amino acid positions 118, 122, 126, and 178 of the amino acid sequence represented by (a) above.

The present invention relates to a complex comprising i) a catalytically inactive site-specific nuclease linked to ii) an array of between two and ten, preferably three to seven effector domains each having a specific chromatin modifying activity, such as, for example, a specific DNA methylation activity, a histone methylation activity, a specific histone acetylation or ubiquitination activity, and/or a specific chromatin demethylation/deacetylation activity, wherein the effector domains are each separated by a linker providing sufficient distance between the domains and the nuclease in order not to substantially interfere with their specific chromatin modifying activities, and the binding of the site-specific nuclease, as well as respective methods involving the complex and use of the complex.

A method for disrupting the formation of circular RNAs in vivo, by disrupting or removing one or more intronic regulatory sequences responsible for circRNA backsplicing. The systems and methods can further be used for inhibiting the expression of therapeutically relevant circular RNAs in vivo related to brain and other disorders, as well as the creation of transgenic animal having one or more circRNA mutations that can be used as models mammalian diseases related to circRNA production and regulation, and in particular human brain disorders.

Aspects of the disclosure relate to methods, compositions, and systems for editing a DNA sequence encoding an endogenous tRNA into a suppressor tRNA using base editing (e.g., to treat a disease caused by a premature termination codon or PTC). Additional aspects relate to compositions comprising a gRNA configured to bind to a DNA sequence encoding an endogenous tRNA. Other aspects relate to complexes comprising a base editor and a gRNA that are capable of editing an endogenous tRNA into a suppressor tRNA. In some aspects, the disclosure further relates to polynucleotides encoding one or more nucleic acid sequences encoding the gRNAs, vectors comprising the polynucleotides, and/or cells comprising the polynucleotides, complexes, gRNAs, and/or vectors disclosed herein. Additional aspects further relate to kits comprising any one of the compositions, complexes, gRNAs, polynucleotides, vectors, and/or cells disclosed herein.

The present disclosure provides recombinant fusogenic proteins comprising a rhabdovirus glycoprotein (G) and a targeting molecule attached to the N-terminus of the rhabdovirus glycoprotein. Further provided are related recombinant polynucleotides, host cells, and pharmaceutical compositions. Recombinant viruses, e.g., recombinant pseudotyped viruses, and cell-derived nanovesicles comprising the recombinant polynucleotides are also provided. Further provided are methods for using the recombinant fusogenic proteins, polynucleotides, viruses, and cell-derived nanovesicles, and/or pharmaceutical compositions thereof, including their use in the treatment of cancer.

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 nucleic acid constructs comprising nucleotide sequences that encode a Cas12a protein. The nucleic acid 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 also provided.

The invention provides a method for increasing the drought resistance of soybean plants by introducing site-specific mutations into stomatal opening genes. The invention also relates to a soybean plant, an isolated part thereof or the seeds thereof, wherein said genes have been inactivated.