The two Gate vectors are in-house-generated Gateway®-compatible pGADT7 and pGBKT7 plasmids of the Matchmaker™ GAL4 Two-Hybrid system (Clontech) for yeast two-hybrid analysis.
Furthermore, upregulated SNF1, HSF1 and wtp53 expression demonstrated that our synthetic AND gate vectors could be activated by combined treatment of radiation and hypoxia.
Construction of AND gate vectors was achieved by conjugation SNF1, HSF1, HSE4 and wtp53 into the downstream of cArG6 promoter in turn.
26, 27 In our experiments, continuous growth inhibition of xenografts with active AND gate genetic circuit demonstrated that after activation, the AND gate vectors could make persistent antitumor effects in vivo and might overcome the temporary expression from inducible promoter.
More importantly, other anti-oncogenes or therapeutic genes can also be conjugated into AND gate vectors to produce targeting antitumor effects such as anti-HER2 gene for breast cancer.
The results were encouraging and indicated that AND gate vectors might have reached into the tumor xenograft cells and were activated by endogenous hypoxia and exogenous radiation and further produced sustained antitumor effect.
The two Gate vectors are in-house-generated Gateway®-compatible pGADT7 and pGBKT7 plasmids of the Matchmaker™ GAL4 Two-Hybrid system (Clontech, Mountain View, CA, USA) for yeast two-hybrid analysis.
Two to 4 h after the 15O-water scans, MRI scans were performed on a 3-T MRI scanner (Achieva, Philips Healthcare) with an 80 mT/m gradient system, using a 32-channel cardiac coil in supine position and retrospectively gated vector ECG for cardiac triggering.
The pTKUbi-gate vector has an expression cassette derived from pTHUbi-Gate 33), pTHUbi-Gate 33 the maize ubiquitin promoter, Gatewhichassette and NOS terminator.
