Shao Yangyang, Lu Ning, Qin Zhongjun, Xue Xiaoli
Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology , Chinese Academy of Sciences , Shanghai 200032 , China.
University of Chinese Academy of Sciences , Beijing 100049 , China.
ACS Synth Biol. 2018 Nov 16;7(11):2706-2708. doi: 10.1021/acssynbio.8b00397. Epub 2018 Nov 5.
Eukaryotic cells usually contain multiple linear chromosomes. Recently, we artificially created a functional single-chromosome yeast via sequential two-chromosome fusion utilizing the high performance of the CRISPR-Cas9 system and homologous recombination in Saccharomyces cerevisiae. In this paper, we adapted this method for the simultaneous fusion of multiple chromosomes. We demonstrated the fusion of two, two-chromosome sets with a 75% positive rate and three-chromosome fusions with a 50% positive rate. We also found that by using an additional selection marker, the positive rate of two-chromosome fusions reached 100%. Due to the simplicity, efficiency, and portability of this method, we expect that it can be easily adapted for multiple-chromosome fusions in other organisms.
真核细胞通常包含多条线性染色体。最近，我们利用酿酒酵母中CRISPR-Cas9系统的高性能和同源重组，通过连续的双染色体融合人工创建了一种功能性单染色体酵母。在本文中，我们采用这种方法实现了多条染色体的同时融合。我们展示了两组双染色体融合的阳性率为75%，三染色体融合的阳性率为50%。我们还发现，通过使用额外的选择标记，双染色体融合的阳性率达到了100%。由于该方法的简单性、高效性和可移植性，我们预计它可以很容易地应用于其他生物体的多染色体融合。
