Disclosed herein are cell genetically engineered cell for AAV production. The genetically engineered cell comprises molecular systems for temporal control of expression of genes required for AAV production. Also disclosed herein are methods of using genetically engineered cells for AAV production.

The invention relates to a polypeptide comprising an amino acid having a norbornene group. Suitably said norbornene group is present as an amino acid residue of a norbornene lysine. The invention also relates to a method of producing a polypeptide comprising a norbornene group, said method comprising genetically incorporating an amino acid comprising a norbornene group into a polypeptide.

Provided herein are compositions and methods for generating polypeptides using non-natural amino acids (nnAAs) and genetic machinery, wherein the modified polypeptides, such as therapeutic polypeptides, bind to albumin, such as serum albumin. Methods of substituting a non-natural amino acid in a first polypeptide to obtain a modified polypeptide, the nnAA in some instances comprising an albumin targeting group, are disclosed, as are methods for making populations of such modified polypeptides. A therapeutic polypeptide, interleukin-1 receptor antagonist (IL-1RA) is exemplified using the disclosed methods.

Methods to generate novel acyl-tRNA species deploy an orthogonal synthetase that accepts -hydroxy acids, -thio acids, N-formyl-L--amino acids, and/or -carboxyl acid monomers (malonic acids) that are formally precursors to polyketide natural products.

Provided herein are, inter alia, non-toxic, bioreactive unnatural amino acids and methods of using same.

The invention relates to genetic incorporation of 2,3-diamino propionic acid (DAP) into polypeptides, to unnatural amino acids comprising DAP, to a tRNA synthetase for charging tRNA with unnatural amino acids comprising DAP, and to methods of using the resulting polypeptides, for example in capturing substrates and/or intermediates in enzymatic reactions. The invention also relates to compounds of formula (I) or (II): ##STR00001##
or salts, solvates, tautomers, isomers or mixtures thereof.

Disclosed herein are methods, compositions and kits for the synthesis of proteins which comprises unnatural amino acids that utilize a mutant tRNA.

The present invention provides in certain embodiments comprising a pyrrolysine-based aminoacyl-tRNA synthetase and a tRNA to form a pyrrolysine-based aminoacyl-tRNA synthetase/tRNA pair. The present invention also provides novel pyrrolysine-based aminoacyl-tRNA synthetases, cells and compositions comprising pyrrolysine-based aminoacyl-tRNA synthetases. The present invention also provides methods of using these pyrrolysine-based aminoacyl-tRNA synthetases.

Disclosed herein are methods, compositions and kits for the synthesis of proteins which comprises unnatural amino acids that utilize a mutant tRNA.

Provided herein are, inter alia, bioreactive unnatural amino acids, compounds containing the unnatural amino acids, and methods of using same.