Posts Tagged ‘Academy’

Generation of a retroviral vector that expresses an anti-HIV-1 tat hammerhead ribozyme.(Clinical report): An article from: Journal of the South Carolina Academy of Science

This digital document is an article from Journal of the South Carolina Academy of Science, published by South Carolina Academy of Science on September 22, 2009. The length of the article is 3271 words. The page length shown above is based on a typical 300-word page. The article is delivered in HTML format and is available immediately after purchase. You can view it with any web browser.

From the author: Ribozymes have emerged as promising therapeutics for HIV and have been shown to bind and cleave target RNA’s in a sequence-specific manner. The HIV-1 regulatory protein tat plays an essential role in the upregulation of viral transcription and elongation of viral transcripts, and has therefore been identified as a target for ribozyme studies. To further study the use of these reagents, we have designed a number of hammerhead ribozymes targeted to specific sequences within the HIV-1 NL43 tat sequence. One of these, Tat5910 and its non-catalytic control Tat5910A, were cloned into the retroviral vector pSuper.retro.neo+GFP (pSRNG) for cell culture studies. Recombinant retrovirus particles were generated by transient transfection of 293T cells using a two-plasmid system consisting of the helper plasmid, pPAM3, and pSRNG5910A. Identical experiments were carried out using two Mo-MLV retroviral vectors, pLNCE, and pLNCLZRz. The resulting recombinant virus was used to transduce NIH-3T3 cells, and virus titer (virus particles/ml) was determined from the number of Green Fluorescent Protein (GFP)- or P-galactosidase-positive cells. Although producer cells transfected with pSRNG5910A expressed GFP, the transfection efficieny was low. This resulted in levels of transduced NIH-3T3 cells that were too low to obtain a reliable titer measurement. Attempts to optimize titer by increasing the number of transfected producer 293T cells was successful using pLNCE and pLNCLZRz. Positive GFP or P-galactosidase expression in NIH-3T3 cells transduced with LNCE and LNCLzRz recombinant virus indicated that our two-plasmid virus production system was successful. Extremely low titer along with lower cellular expression of GFP suggests that the pSRNG5910 vectors may be better suited to virus production using a stable producer cell line such as PA317

Citation Details
Title: Generation of a retroviral vector that expresses an anti-HIV-1 tat hammerhead ribozyme.(Clinical report)
Author: Lindsey E. Padgett
Publication: Journal of the South Carolina Academy of Science (Magazine/Journal)
Date: September 22, 2009
Publisher: South Carolina Academy of Science
Volume: 7 Issue: 2 Page: 18(5)

Article Type: Clinical report

Distributed by Gale, a part of Cengage Learning

List Price: $ 9.95

Price: $ 9.95