In vivo transduction of HIV-1-derived lentiviral particles engineered for macrolide-adjustable transgene expression

Abstract

Macrolide-adjustable lentivectors enable robust and precise in vitro and in vivo transgene fine-tuning which may give future gene therapy trials a new impetus.

Poly(ADP-Ribose) Polymerase 1 Promotes Transcriptional Repression of Integrated Retroviruses

“…These cells efficiently support the early postentry steps of the gammaretrovirus and lentivirus life cycle (35)(36)(37)(38)(39). PARP-1 is the only member of the PARP family with catalytic activity in these cells, and DT40 cells are viable after PARP-1 knockout (34).…”

“…In addition, chicken cells can be used as a reliable model to study the molecular events that occur from entry to proviral transgene expression characteristic of the gammaretrovirus and lentivirus life cycle. Chicken cells are highly permissive to transduction with singleround infection by murine leukemia virus (MLV) and HIV reporter viruses (35)(36)(37)(38)(39), suggesting that the molecular mechanisms required for uncoating, reverse transcription, DNA integration, and transgene expression of these retroviruses are conserved in these cells. Further evidence supporting the conservation of the molecular mechanisms implicated in the early steps of HIV-1 infection in chicken cells indicates that, as in human cells, HIV integrates preferentially inside active transcription units in the chicken genome (35).…”

Modulation of Cardiomyocyte Electrical Properties Using Regulated Bone Morphogenetic Protein-2 Expression

“…16 Based on the ability of pseudotyped lentiviral vectors to transduce a wide variety of cell types and tissues, including quiescent cells, transgenic lentiviral particles have become an attractive gene-transfer tool for cardiac cells. [17][18][19] Moreover, lentiviral vectors have recently been engineered to regulate transgene expression, [20][21][22] thus providing a powerful tool for functional genomic research; 23 production of transgenes in animal models of human diseases; 24 in vivo or ex vivo titration of pharmaceutical proteins within a therapeutic range; [25][26][27] and rational reprogramming of the proliferation, differentiation, or apoptosis pathways to engineer specific cell and tissue phenotypes. 20,[28][29][30][31][32] Capitalizing on the potential of inducible viral transgene expression 33 and on recent advances in novel regulation systems, 34 we selected two lentivirus-based generegulation systems to genetically manipulate neonatal cardiomyocytes: a streptogramin-responsive expression system, 20,35 in which addition of pristinamycin I (PI) switches off transgene expression (OFF system), and a gas-inducible expression system, in which the presence of administered gaseous acetaldehyde (AcAl ) switches on transgene expression (ON system).…”

InXy and SeXy, compact heterologous reporter proteins for mammalian cells

“…Secreted mammalian reporter proteins have been used extensively for functional genomic research (Kramer and Fussenegger, 2005), as marker genes in vivo (Mitta et al, 2005), for prototype gene therapy scenarios (Liu et al, 2004;Weber et al, 2004) as well as for high-throughput screening initiatives (Durocher et al, 2000;Lee et al, 2004). A variety of intracellular reporter proteins are available for use in mammalian cells such as (i) specific enzymes (bgalactosidase, thymidine kinase or chloramphenicol acetyltransferase) Gorman et al, 1982;Mannhaupt et al, 1988;Searle et al, 1985), (ii) fluorescent reporter proteins represented by the pioneering green fluorescent protein (e.g.…”