Gene Delivery to Mammalian Cells Vol 2: Viral Gene Transfer by William C. Heiser

By William C. Heiser

Hugely skilled researchers describe in step by step aspect tools that experience confirmed most valuable in supplying genes to mammalian cells. quantity 2: Viral Gene move concepts, information techniques for offering genes to cells in vitro and in vivo, together with using lentiviral vectors. An accompanying quantity, quantity 1: Nonviral Gene move options specializes in gene supply by means of numerous chemical and actual equipment, together with ultrasound, biolistics, peptides, PNA clamps, liposomes, microinjection, electroporation, particle bombardment, dendrimers, and hydrodynamics.

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Weber, J. , and Massie, B. (2000) Proteasedeleted adenovirus vectors and complementing cell lines: potential applications of single-round replication mutants for vaccination and gene therapy. Hum. Gene Ther. 11, 1341–1353. 16. Greber, U. , and Helenius, A. (1996) The role of the adenovirus protease on virus entry into cells. EMBO J. 15, 1766–1777. 17. Elahi, S. , and Massie, B. (2002) Adenovirus-based libraries: efficient generation of recombinant adenoviruses by positive selection with the adenovirus protease.

1998) A simplified system for generating recombinant adenoviruses. Proc. Natl. Acad. Sci. USA 95, 2509–2514. 13. , and Hieter, P. (1994) Efficient manipulation of the human adenovirus genome as an infectious yeast artificial chromosome clone. Proc. Natl. Acad. Sci. USA 91, 6186–6190. 14. Mizuguchi, H. and Kay, M. A. (1998) Efficient. construction of a recombinant adenovirus vector by an improved in vitro ligation method. Hum. Gene Ther. 9, 2577–2583. DNA Delivery to Cells in Culture 27 15. , Weber, J.

1% (w/v) trypsin-EDTA in HBSS. 6. After the enzymatic dissociation, centrifuge and resuspend the isolated muscle cells in growth medium. 7. Preplate the cells on collagen-coated 6-well plates and incubate at 37°C in 5% CO2. After about 1 h, decant the supernatant containing nonadherent cells from the well and replate in a fresh collagen-coated well (see Note 1). 8. Subculture myoblasts before they become confluent, to avoid differentiation into myotubes. 2. Differentiation of Myoblasts into Myotubes 1.

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