Dataset Information
New ?- and SIN ?-retrovectors for safe transduction and specific transgene expression in pancreatic ? cell lines.
Ontology highlight
ABSTRACT: Background
Viral vectors are invaluable tools to transfer genes and/or regulatory sequences into differentiated cells such as pancreatic cells. To date, several kinds of viral vectors have been used to transduce different pancreatic cell types, including insulin-producing ? cells. However, few studies have used vectors derived from « simple » retroviruses, such as avian ?- or mouse ?-retroviruses, despite their high experimental convenience. Moreover, such vectors were never designed to specifically target transgene expression into ? cells.Results
We here describe two novel ?- or SIN (Self-Inactivating) ?-retrovectors containing the RIP (Rat Insulin Promoter) as internal promoter. These two retrovectors are easily produced in standard BSL2 conditions, rapidly concentrated if needed, and harbor a large multiple cloning site. For the SIN ?-retrovector, either the VSV-G (pantropic) or the retroviral ecotropic (rodent specific) envelope was used. For the ?-retrovector, we used the A type envelope, as its receptor, termed TVA, is only naturally present in avian cells and can efficiently be provided to mammalian ? cells through either exogenous expression upon cDNA transfer or gesicle-mediated delivery of the protein. As expected, the transgenes cloned into the two RIP-containing retrovectors displayed a strong preferential expression in ? over non-? cells compared to transgenes cloned in their non-RIP (CMV- or LTR-) regulated counterparts. We further show that RIP activity of both retrovectors mirrored fluctuations affecting endogenous INSULIN gene expression in human ? cells. Finally, both ?- and SIN ?-retrovectors were extremely poorly mobilized by the BXV1 xenotropic retrovirus, a common invader of human cells grown in immunodeficient mice, and, most notably, of human ? cell lines.Conclusion
Our novel ?- and SIN ?-retrovectors are safe and convenient tools to stably and specifically express transgene(s) in mammalian ? cells. Moreover, they both reproduce some regulatory patterns affecting INSULIN gene expression. Thus, they provide a helpful tool to both study the genetic control of ? cell function and monitor changes in their differentiation status.
SUBMITTER: Albagli O
PROVIDER: S-EPMC6580483 | biostudies-literature | 2019 Jun
REPOSITORIES: biostudies-literature
Publications
New α- and SIN γ-retrovectors for safe transduction and specific transgene expression in pancreatic β cell lines.
BMC biotechnology 20190617 1
<h4>Background</h4>Viral vectors are invaluable tools to transfer genes and/or regulatory sequences into differentiated cells such as pancreatic cells. To date, several kinds of viral vectors have been used to transduce different pancreatic cell types, including insulin-producing β cells. However, few studies have used vectors derived from « simple » retroviruses, such as avian α- or mouse γ-retroviruses, despite their high experimental convenience. Moreover, such vectors were never designed to ...[more]
