A woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) and cPPT is included in each vector to increase gene expression levels in transduced cells. This ensures that, after integration in the target cell genome, only the internal promoter of interest is active and no further viral replication or infection occurs. The engineered virus is replication incompetent and has a self-inactivating viral promoter (deleted U3). To summarize, the third generation lentiviral vectors employed in our studies are typically pseudotyped with a VSV-G envelope that allows stability and tropism for cells of multiple species including mice. We have previously published a detailed description of the lentiviral backbone and technique for packaging high titer virus for in vivo studies, and several of these protocols may be downloaded here by clicking on protocols. Kotton worked with the original pHAGE backbone during his postdoctoral fellowship in the Mulligan laboratory and the backbone has since been adapted to express the transgenes indicated on this page. The lentiviral system we employ is based on an HIV-1-based backbone, named ‘pHAGE’ (standing for plasmid HIV-1 Alex Gustavo George Enhanced) originally developed in the laboratory of Dr. PHAGE-EF1aL-TagBFP-W W=WPRE sequence L=long version of promoter Background on pHAGE: A third generation lentiviral vector These and other vectors are also available to Boston University Investigators at our CReM vector core webpage, please visit here for a more complete inventory.įor information on our iPS cell bank, please visit our CReM iPS Cell Core website. Kotton, serves on the Addgene Board of Directors. Some of our vectors are also available from Addgene, where our PI, Dr. pdf files by clicking on the relevant link below for each vector.
#Vector nti express import sequencing for free#
Vector maps and sequences are available for free download as genbank formatted. Please visit our PROTOCOLS page for all protocols, including ESC/iPSC lung differentiation, lentiviral packaging and titering, or intratracheal lentiviral transduction of lung macrophages! Or take our annual hands-on 1 week course! Sign up through the CReM iPSC Core webpage. Requesting our cells is easy, click here for details!ĭifferentiating iPSCs in your own lab is a little harder, but we’ll show you how: Vectors, Protocols, and Cells So many cells, so little time….
0 kommentar(er)
