Clones of hAFSC were transduced with a lentiviral vector (SMPU-R-MNCU3-LUC based on HIV-1 that transduces the firefly luciferase gene) made by the Vector Core Facility at Childrens Hospital Los Angeles following standard protocols as reported in the paper by Crooks et al [29].
Stably transduced HCT116 tumor cells expressing the GFP-LC3 gene were generated by lentiviral transfection of the pCT-autophagosome-GFP Vector (Lentiviral Laboratory of the Vector Core Facility of the University of Pittsburgh) and selected with puromycin.
An AAV-2 vector expressing GFP from a chicken b-actin promoter (AAVCAGGFP) was purchased from the vector core facility.
The F3 families, their parents, and control lines were evaluated for resistance to RHBV and feeding damage caused by the vector insect in the greenhouse facilities at the CIAT headquarters in Palmira, Colombia.
Two more variables, number of doctors and nurses in each health facility, were also added making the number of variables in the vector of health facility characteristics seven.
The AAVCAGsCD59 virus was produced and purified by the UMass Vector Core Facility, using AAV serotype 2 capsid proteins.
AAV-EF1a-DIO-mCherry (Anthony et al., 2014; Lee et al., 2014) was constructed in house (Dr Todd Anthony) and packaged by the University of Pennsylvania vector core facility.
All lentiviruses were produced by the Viral Vectors Core Facility of the Hope Center for Neurological Diseases at Washington University School of Medicine.
The primers used to amplify the clones are summarized in Table 5. PCR products of interest were cloned into pCR®2.1-TOPO vector and sequenced by the NAPS facility at the University of British Columbia.
AAV vectors were generated by the University of Pennsylvania Vector Core facility by triple transfection of subconfluent HEK293 cells using 3 plasmids: an AAV trans-plasmid encoding AAV2 capsid, an adenovirus helper plasmid pΔF6, and an AAV cis-shuttle plasmid expressing eGFP driven by a CMV promoter (pENN.AAV.CMV.PI.eGFP.WPRE.bGH).
PCR products of the VRN-B1 gene were purified with the Qiagen spin miniprep kit (Qiagen Sciences Inc ., cloned into the pCR2.1 TOPO vector (Invitrogen Inc., Carlsbad, CA), and then sequenced by the DNA Facility at Iowa State University (Ames, Iowa).
