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In 2009, the European Molecular Biology Organization released the report "Making the Most of Synthetic Biology Strategies for Synthetic Biology Development in Europe" to introduce a ____________________ for European synthetic biology research.

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To enable efficient genomic manipulation, Church and colleagues at Harvard Medical School developed a platform called multiplex automated genome engineering (MAGE) in 2011, which has been used to rapidly alter multiple loci in the E. coli genome, including the _____________________________ of all TAG stop codons with the synonymous TAA codon.

RNA-based circuit engineering also underwent an expansion during 2008-2013, as biosensing functions gave way to RNA-based computation. RNA devices were built to control the regulatory logic of gene expression, and RNA design tools were developed to enable the precise, predictable control of heterologous and ____________________________.

The CRISPR–Cas immunity system in bacteria and archaea was re-purposed to enable genome-wide transcriptional control. Type II CRISPR–Cas systems use RNA-directed DNA binding by the nuclease Cas9 to detect and cleave invading bacteriophage and other __________________________, and independent groups developed Cas9 nuclease mutants that enable RNA-directed DNA binding by Cas9 without subsequent DNA cleavage.

The report "Synthetic Biology" was submitted to the US Congress by the US Department of Energy in ______. Three technical challenges in the synthetic-biology field were formulated: i) Genome scale engineering tools, DNA synthesis and assembly, and analytical tools; ii) Biological design principles, genetically tractable organisms/chassis, a minimal cell and in vitro systems, tools for plant systems, and biocontainment mechanisms; and iii) Computational tools, information standards, and databases.

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