Abstract: The present invention relates to recombinant adeno-associated viruses (rAAVs) having capsid proteins engineered to be a tropic or have limited tropism which may be further engineered by insertion of a targeting domains that confer and/or enhance desired properties, particularly increased transduction in CNS or muscle cells or other target tissue relative to a rAAV having a reference capsid.

Abstract: The present invention relates to recombinant adeno-associated viruses (rAAVs) having capsid proteins that have a tropism for ocular tissue. The rAAVs have capsids that have enhanced or increased transduction of ocular tissues as compared to reference rAAVs. Such rAAVs may be useful in delivering transgenes encoding therapeutic proteins for the treatment of ocular disease.

Abstract: The present invention relates to recombinant adeno-associated viruses (rAAVs) having capsid proteins engineered to include amino acid sequences and/or amino acid substitutions that confer and/or enhance desired properties, particularly increased transduction in CNS or muscle cells relative to a rAAV having a reference capsid.

Abstract: The present invention relates to recombinant adeno-associated viruses (rAAVs) having capsid proteins engineered to include amino acid sequences that confer and/or enhance desired properties. In particular, the invention provides engineered capsid proteins comprising peptide insertions from heterologous proteins inserted within or near variable region IV (VR-IV) of the virus capsid, such that the insertion is surface exposed on the AAV particle. The invention also provides capsid proteins that direct rAAVs to target tissues, in particular, capsid proteins comprising peptides derived from erythropoietin or dynein that are inserted into surface-exposed variable regions and that target rAAVs to retinal tissue and/or neural tissue, including the central nervous system, and deliver therapeutics for treating neurological and/or eye disorders.

Abstract: The invention in at least one embodiment includes a method to convert a fluorescent protein into a modified fluorescent protein in a monomeric state where a dimerization interface is identified in a template protein in a homodimer state. One or more targeted mutation(s) are introduced at the dimerization interface to disrupt dimerization and favor the monomeric state. In another embodiment of the invention, one or more chromophore binding domains are identified in a template protein. One or more targeted mutations are introduced in the chromophore binding domain(s) to change the configuration of a biliverdin chromophore in the template protein from a 15Z anti configuration to a 15E anti configuration.

Abstract: Methods for the creation of one or more user-defined mutations that can be located anywhere in a target sequence, such as in a gene are disclosed. These methods can be used on a single-stranded or double-stranded template and, for a single-stranded template, generally include: (a) conducting a first amplification reaction in the presence of a thermostable DNA polymerase and a thermostable DNA ligase to synthesize a mutagenized strand of DNA comprising at least one mutagenic oligonucleotide relative to a complementary single-stranded template DNA comprising a target nucleic acid molecule in a circular DNA vector; (b) conducting a second amplification reaction in the presence of a thermostable DNA polymerase and a thermostable DNA ligase to synthesize a complementary mutant strand of DNA; and (c) degrading the template DNA and non-covalently closed circular nucleic acid molecules to obtain a mutation-containing double-stranded DNA product.

Abstract: The presently disclosed subject matter provides methods for the creation of one or more user-defined mutations that can be located anywhere in a target sequence, such as in a gene. These mutations can comprise single mutations, multiple mutations, or a comprehensive codon mutagenesis library, in which all possible single codon substitutions in a gene are created. These methods can be used on a single-stranded or double-stranded template.

Abstract: The presently disclosed subject matter relates to modified Kunkel mutagenesis methods that use a thermostable DNA polymerase and a thermostable DNA ligase.

Abstract: The presently disclosed subject matter relates to modified Kunkel mutagenesis methods that use a thermostable DNA polymerase and a thermostable DNA ligase.

Abstract: The presently disclosed subject matter provides methods for the creation of one or more user-defined mutations that can be located anywhere in a target sequence, such as in a gene. These mutations can comprise single mutations, multiple mutations, or a comprehensive codon mutagenesis library, in which all possible single codon substitutions in a gene are created. These methods can be used on a single-stranded or double-stranded template.

Abstract: The presently disclosed subject matter provides methods for the creation of one or more user-defined mutations that can be located anywhere in a target sequence, such as in a gene. These mutations can comprise single mutations, multiple mutations, or a comprehensive codon mutagenesis library, in which all possible single codon substitutions in a gene are created. These methods can be used on a single-stranded or double-stranded template.