This invention relates to site-specific integration and expression of recombinant proteins in eukaryotic cells. In particular, the invention includes compositions and methods for improved expression of antibodies including bispecifc antibodies in eukaryotic cells, particularly Chinese hamster (Cricetulus griseus) cell lines, by employing an expression-enhancing locus.

A mutant SaCas9 protein such as a protein having an amino acid sequence resulting from mutations of glutamic acid at the 782-position to lysine (E782K), leucine at the 800-position to arginine (L800R), asparagine at the 968-position to arginine (N968R), asparagine at the 985-position to alanine (N985A), arginine at the 991-position to alanine (R991A), alanine at the 1021-position to serine (A1021S), threonine at the 927-position to lysine (T927K), lysine at the 929-position to asparagine (K929N), and isoleucine at the 1017-position to phenylalanine (I1017F) in SEQ ID NO: 2 has relaxed restriction on target sequence while maintaining binding ability to guide RNA, and is useful as a tool for gene editing.

The present disclosure is in the technical field of synthetic biology and metabolic engineering. The disclosure provides engineered viable bacteria. In particular, the disclosure provides viable bacteria with reduced cell wall biosynthesis additionally modified for production of glycosylated product. The disclosure further provides methods of generating viable bacteria and uses thereof. Furthermore, the disclosure in the technical field of fermentation of metabolically engineered microorganisms producing glycosylated product.

Provided herein are compositions that include at least two different nucleic acid vectors, where each of the at least two different vectors includes a coding sequence that encodes a different portion of an otoferlin protein, and the use of these compositions to treat hearing loss in a subject.

Methods and compositions are provided for modifying the genome of Bacillus sp. cells without the use of a selectable marker and without the use of a guided Cas endonuclease. The disclosure includes methods for integrating donor DNA sequences into the genome of a Bacillus sp. cell without the use of a selectable marker and without the use of Cas endonucleases into said genome, as well as methods for deleting genes of interest and/or providing point mutations into the genome of Bacillus sp. cells.

The present disclosure relates to Salmonella Gallinarum mutant strains and uses thereof. A vaccine composition according to an aspect has no risk of recovering pathogenicity, has no residual pathogenicity due to detoxification of an endotoxin, and does not cause lesions and bacterial re-isolation, thereby exhibiting significantly improved safety compared to the existing fowl typhoid vaccines. In addition, since the vaccine composition induces a high-level immune response even when administered to young chicks, it may be used regardless of age, and as the vaccine strain may be used as a live vaccine having an excellent protective capability by itself, the vaccine composition may be useful for preventing and alleviating fowl typhoid.

A transkingdom platform for the delivery of therapeutics to target cells. The system maintains the export and uptake functions of Inv while modifying its targeting away from 1 integrin to other proteins expressed on the surface of target eukaryotic cells (i.e., a cell surface protein) or chemical moieties (i.e., a cell surface chemical moiety) expressed on the surface of a target eukaryotic cell by replacing D4 and D5 of Inv with a binding domain from a heterologous protein via genetic engineering. These heterologous proteins could be derived from bacterial, fungal, animal, or viral genomes. This engineering would result in the construction of a chimeric Inv protein in which D1-D3 (i.e., the non-binding domains) are fused in frame to an alternative binding domain derived from a heterologous protein. The alternative binding domain would interact with a different cell surface protein or chemical moiety, which can in some instances be referred to as a receptor, on the surface on the surface of a eukaryotic cell, thereby allowing specific targeting to cells independent of Inv's intrinsic 1 integrin binding.

The disclosure relates to insecticidal toxin resistance management and screening of novel insecticidal toxins. One embodiment relates to the isolation, characterization, compositions, and methods of use relating to polynucleotides encoding novel insecticidal toxin receptors and the polypeptides encoded thereby. The polynucleotides and polypeptides are useful in identifying or designing novel insecticidal toxin receptor ligands including novel insecticidal toxins.

The present invention discloses base editing systems for mutating a base C to A and a base C to G and applications thereof. The base editing system for mutating C to A disclosed in the present invention includes cytosine deaminase AID and nCas9 nuclease or includes cytosine deaminase AID, nCas9 nuclease and uracil DNA glycosidase; the base editing system for mutating C to G of the present invention includes cytosine deaminase APOBEC, nCas9 nuclease and uracil DNA glycosidase. The experiments show that a combination of the three base editing systems for mutating C to A, C to T and A to G can realize a mutation of A, T, C or G to any base in both prokaryotes and eukaryotes.

Provided herein, in some aspects, are tools (e.g., methods, compositions and nucleic acids) for building genetic circuits in Bacteroides and Parabacteroides bacteria, as well as the bacteria containing the genetic circuits.