RESUMO
We present a deterministic workflow for genotyping single and double transgenic individuals directly upon nascence that prevents overproduction and reduces wasted animals by two-thirds. In our vector concepts, transgenes are accompanied by two of four clearly distinguishable transformation markers that are embedded in interweaved, but incompatible Lox site pairs. Following Cre-mediated recombination, the genotypes of single and double transgenic individuals were successfully identified by specific marker combinations in 461 scorings.
Assuntos
Animais Geneticamente Modificados , Engenharia Genética/métodos , Vetores Genéticos/metabolismo , Técnicas de Genotipagem , Integrases/genética , Tribolium/genética , Animais , Embrião não Mamífero , Feminino , Corantes Fluorescentes/química , Corantes Fluorescentes/metabolismo , Expressão Gênica , Engenharia Genética/economia , Marcadores Genéticos , Vetores Genéticos/química , Heterozigoto , Histonas/genética , Histonas/metabolismo , Homozigoto , Integrases/metabolismo , Masculino , Microscopia de Fluorescência , Regiões Promotoras Genéticas , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismoRESUMO
Launching a new drug on the market is an extremely time-consuming and expensive process. The total costs from the lab bench to the patient's bedside are in the range of $800 million for each new compound. Innovative pre-clinical assays are urgently needed to select the most promising drug candidates. High-throughput molecular screening does not provide information on the effects on cellular functions. Testing on animals is expensive, ethically controversial, and poorly predictive of the response in humans. Conventional two-dimensional (2D) cellular assays do not accurately reflect the drug response in vivo. To overcome these limitations, biotechnologists are developing three-dimensional (3D) cultures. 3D cultures provide more accurate compound screening and can eliminate toxic and ineffective substances at an early stage. Moreover, 3D cultures can accomplish the 3R agenda (refinement, reduction, and replacement) for the replacement of toxicity testing on animals. We provide an up-to-date overview on the patents in the field.
