PhiC31 integrase mediates integration in cultured synovial cells and enhances gene expression in rabbit joints

Abstract

The ability of phiC31 integrase to facilitate genomic integration in synovial cells and increase transgene expression in the rabbit synovium suggests that, in combination with more efficient DNA delivery methods, this integrase system could be beneficial for treatment of rheumatoid arthritis and other joint disorders.

Long-term transgene expression in mouse neural progenitor cells modified with ϕC31 integrase

“…We observed nearly 40-fold higher levels of luciferase expression in the mNPCs modified with ϕC31 integrase as compared to those without at 8 weeks after nucleofection. The stable expression of the transgene has been attributed to the ability of ϕC31 integrase to mediate integration of the gene into host chromosomes (Groth et al, 2000;Thyagarajan et al, 2001;Olivares et al, 2002;Ortiz-Urda et al, 2002;Quenneville et al, 2004;Chalberg et al, 2005;Ishikawa et al, 2006;Bertoni et al, 2006;Keravala et al, 2006;Portlock et al, 2006). We obtained PCR evidence for integration of the luciferase plasmid at mpsL1, a known genomic hot-spot utilized by ϕC31 integrase (Olivares et al, 2002;Bertoni et al, 2006;Portlock et al, 2006).…”

ΦC31 Integrase Mediates Efficient Site-Specific Integration in Sheep Fibroblasts

“…Pseudo-attP sites have traits similar to those of human, mouse, rat, and Drosophila. 5,7,10) By using the ÈC31 integrase gene delivering system, elevated integration frequency and effective gene expression can be effected in sheep fibroblasts. Furthermore, we have obtained large numbers of integrase-mediated transgenic embryos by somatic-cell nuclear transfer (data not shown).…”

“…These have been identified in the genome of human, mouse, rat, rabbit, and Drosophila. [6][7][8][9] Furthermore, unlike the widely used Cre and FLP recombinases, which can catalyze both integration and excision, ÈC31 integrase-mediated integration is unidirectional. This makes the ÈC31 integrase system preferable for transgene insertion.…”

PhiC31 Integrase Induces Efficient Site-Specific Recombination in the Capra hircus Genome

“…According to the published data (Groth et al, 2004;Chalberg et al, 2005Chalberg et al, , 2006Keravala et al, 2006;Ma et al, 2006;Ehrhardt et al, 2007; Qu et al, 2012), the sequences we retrieved around the pseudo attP sites were about 200 bp. As a control, 97 random sequences corresponding to these species were similarly analyzed by motif finder software.…”

“…It has been reported that the integrations induced by fC31 integrase support high-level and long-term expression of foreign genes (Ortiz-Urda et al, 2002Quenneville et al, 2004;Held et al, 2005;Keravala et al, 2006;Ou et al, 2009). We observed long-term expression up to 30 days for GFP integrated at the pseudo attP sites of C. hircus fibroblast cells, Figure 3A.…”