Synthetic Saccharomyces cerevisiae Genome sequencing of synIXR and semi-synVIL. Saccharomyces cerevisiae

Synthetic chromosome rearrangement and modification by loxP-mediated evolution (SCRaMbLE) generates combinatorial genomic diversity: recombinase sites are encoded in a synthetic chromosome, and tightly controlled induction of a modified recombinase creates a population of targeted structural variations. We used this method to scramble large regions of two yeast chromosomes: yeast synthetic chromosome arm synIXR, with 43 recombinase target sites, and synthetic region synVIL, with 5 recombinase sites. Deconvolving the scrambled genomes provides insight into genome structure and function and reveals whether off-target activity or ectopic recombinations interfere with the synthetic design. Mutant; synthetic chromosome rearrangement and modification by loxP-mediated evolution (SCRaMbLE).

基于loxP介导进化的人工染色体重排与修饰（Synthetic chromosome rearrangement and modification by loxP-mediated evolution, SCRaMbLE）可产生组合式基因组多样性：该技术将重组酶靶位点编码于人工染色体中，通过严格调控修饰型重组酶的诱导表达，可构建出携带靶向结构变异的细胞群体。本研究利用该方法对两条酵母染色体的大片段区域进行重排：分别为携带43个重组酶靶位点的酵母人工染色体臂synIXR，以及携带5个重组酶靶位点的人工区域synVIL。对重排基因组的解析，可为基因组结构与功能研究提供深入见解，并可揭示脱靶活性或异位重组是否会干扰人工设计的基因组方案。突变体；基于loxP介导进化的人工染色体重排与修饰（SCRaMbLE）。