Development of Lentiviral Vectors for Targeted Integration and Protein Delivery.
The method in this chapter describes the design of human immunodeficiency virus type 1 (HIV-1) integrase (IN)-fusion proteins which we have developed to transport different proteins into the nuclei of lentiviral vector (LV)-transduced cells. The IN-fusion protein cDNA is incorporated into the LV packaging plasmid, which leads to its incorporation into vector particles as part of a large Gag-Pol polyprotein. This specific feature of protein packaging enables also the incorporation of cytotoxic and proapoptotic proteins, such as frequently cutting endonucleases and P53. The vectors can hence be used for various protein transduction needs. An outline of the necessary methods is also given to study the functionality of a chosen IN-fusion protein in a cell culture assay.
Department of Biotechnology and Molecular Medicine, A.I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland.
Diana
D
Schenkwein
Department of Biotechnology and Molecular Medicine, A.I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland. seppo.ylaherttuala@uef.fi.
Seppo
S
Ylä-Herttuala
D005822:Genetic Vectors; D019427:HIV Integrase / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection / Q000379:methods*
D011993:Recombinant Fusion Proteins; D019427:HIV Integrase
Genomic safe harbor; Integrase fusion protein; Protein transduction; Targeted integration; Vector production