Transduction of non-adherent bone marrow cells using ViroMag R/L

Non-adherent bone marrow cells were transduced by shRNA lentiviral particles using ViroMag R/L.

This paper shows the efficiency of ViroMag R/L from OZ Biosciences to transduce bone marrow cells with lentiviral particles encoding for shRNA.

article reference: J Bone Miner Res. 2013 Feb 20. doi: 10.1002/jbmr.1884.

Role of ATF7-TAF12 interactions in the VDR hyper-sensitivity of osteoclast precursors in Paget's disease.

Teramachi J, Hiruma Y, Ishizuka S, Ishizuka H, Brown JP, Michou L, Cao H, Galson DL, Subler MA, Zhou H, Dempster DW, Windle JJ, Roodman GD, Kurihara N.

Abstract

Osteoclast (OCL) precursors from many Paget's disease (PD) patients express measles virus nucleocapsid protein (MVNP) and are hypersensitive to 1,25-(OH)(2) D(3) . The increased 1,25-(OH)(2) D(3) sensitivity is mediated by TAF12, a co-activator of VDR, which is present at much higher levels in MVNP-expressing OCL precursors than normals. These results suggest that TAF12 plays an important role in the abnormal OCL activity in PD. However, the molecular mechanisms underlying both 1,25-(OH)(2) D(3) 's effects on OCL formation and the contribution of TAF12 to these effects in both normals and PD patients are unclear. Inhibition of TAF12 with a specific TAF12 antisense construct decreased OCL formation and OCL precursors sensitivity to 1,25-(OH)(2) D(3) in PD patient bone marrow samples. Further, OCL-precursors from transgenic mice in which TAF12 expression was targeted to the OCL lineage (TRAP-TAF12 mice), formed OCL at very low levels of 1,25-(OH)(2) D(3) , although the OCL failed to exhibit other hallmarks of PD OCL, including RANKL hyper-sensitivity and hyper-multinucleation. ChIP analysis of OCL precursors using an anti-TAF12 antibody demonstrated that TAF12 binds the 24-hydroxylase (CYP24A1) promoter, which contains two functional vitamin D response elements (VDRE), in the presence of 1,25-(OH)(2) D(3) . Since TAF12 directly interacts with the ATF7 transcription factor and potentiates ATF7-induced transcriptional activation of ATF7-driven genes in other cell types, we determined if TAF12 is a functional partner of ATF7 in OCL precursors. Immunoprecipitation of lysates from either WT or MVNP-expressing OCL with an anti-TAF12 antibody followed by blotting with an anti-ATF7 antibody, or vice versa, showed that TAF12 and ATF7 physically interact in OCL. Knockdown of ATF7 in MVNP-expressing cells decreased CYP24A1 induction by 1,25-(OH)(2) D(3) as well as TAF12 binding to the CYP24A1 promoter. These results show that ATF7 interacts with TAF12 and contributes to the hyper-sensitivity of OCL precursors to 1,25-(OH)(2) D(3) in PD. © 2013 American Society for Bone and Mineral Research.

ViroMag R/L from OZ Biosciences is a magnetic nanoparticles formulation optimized for increasing any kind of virus infection and transduction both in vitro and in vivo.