Gene transfer to synovial fibroblast: methods and evaluation in the SCID mouse model.
The use of gene transfer techniques has become of utmost importance both for the analysis of molecular pathways of rheumatic joint destruction and for the evaluation of novel therapeutic concepts to treat rheumatic diseases. However, gene transfer into synovial fibroblasts faces several challenges, which result mainly from the lack of specific surface markers and the low-proliferation rate of these cells. This chapter describes both nonviral and viral strategies of transferring gene constructs into synovial fibroblasts. It focuses on the use of lipofection for the gene transfer of siRNA to synovial fibroblasts and the use of AMAXA-nucleofection for the nonviral transfer of gene expression constructs. In addition, retro- and lentiviral strategies of gene transfer are introduced. Finally, the SCID mouse in vivo model of rheumatoid joint destruction is described as a means of evaluating the effects of gene transfer on the invasiveness of synovial fibroblasts.
Meinecke
Ingmar
I
Rutkauskaite
Edita
E
Cinski
Antje
A
Müller-Ladner
Ulf
U
Gay
Steffen
S
Pap
Thomas
T
D000818:Animals; D001169:Arthritis, Experimental / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D001483:Base Sequence; D018274:Electroporation / Q000379:methods; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D013583:Synovial Membrane / Q000378:metabolism*; D019065:Virus Assembly
D034741:RNA, Small Interfering