Efficient viral delivery system for unnatural amino acid mutagenesis in mammalian cells.
Proc Natl Acad Sci U S A
; 110(29): 11803-8, 2013 Jul 16.
Article
en En
| MEDLINE
| ID: mdl-23818609
ABSTRACT
Here we report the development of a baculovirus-based delivery system that enables the efficient incorporation of unnatural amino acids into proteins in mammalian cells. We have exploited the large cargo-capacity (>30 kb) and stability of the double-stranded DNA genome of baculovirus to deliver to a variety of cell types all of the components required to genetically incorporate novel amino acids. These include the engineered tRNA/aminoacyl-tRNA synthetase pair and the nonsense mutant of the target gene. Mammalian cell transduction efficiency of baculovirus was significantly improved by incorporating genetic elements from mammalian viruses. Two polyspecific tRNA/aminoacyl-tRNA synthetase pairs were inserted into this expression system, enabling the site-specific incorporation of a variety of unnatural amino acids with novel chemical and biological properties into proteins.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
ADN Viral
/
ARN de Transferencia
/
Mutagénesis
/
Sistemas de Liberación de Medicamentos
/
Aminoácidos
/
Aminoacil-ARNt Sintetasas
Límite:
Animals
Idioma:
En
Revista:
Proc Natl Acad Sci U S A
Año:
2013
Tipo del documento:
Article
País de afiliación:
Estados Unidos