Xinzhi Zou

All Publications

• Engineering the ribosomal DNA in a megabase synthetic chromosome SCIENCE Zhang, W., Zhao, G., Luo, Z., Lin, Y., Wang, L., Guo, Y., Wang, A., Jiang, S., Jiang, Q., Gong, J., Wang, Y., Hou, S., Huang, J., Li, T., Qin, Y., Dong, J., Qin, Q., Zhang, J., Zou, X., He, X., Zhao, L., Xiao, Y., Xu, M., Cheng, E., Huang, N., Zhou, T., Shen, Y., Walker, R., Luo, Y., Kuang, Z., Mitchell, L. A., Yang, K., Richardson, S. M., Wu, Y., Li, B., Yuan, Y., Yang, H., Lin, J., Chen, G., Wu, Q., Bader, J. S., Cai, Y., Boeke, J. D., Dai, J. 2017; 355 (6329)

  Abstract

  We designed and synthesized a 976,067-base pair linear chromosome, synXII, based on native chromosome XII in Saccharomyces cerevisiae SynXII was assembled using a two-step method, specified by successive megachunk integration and meiotic recombination-mediated assembly, producing a functional chromosome in S. cerevisiae. Minor growth defect "bugs" detected in synXII, caused by deletion of tRNA genes, were rescued by introducing an ectopic copy of a single tRNA gene. The ribosomal gene cluster (rDNA) on synXII was left intact during the assembly process and subsequently replaced by a modified rDNA unit used to regenerate rDNA at three distinct chromosomal locations. The signature sequences within rDNA, which can be used to determine species identity, were swapped to generate a Saccharomyces synXII strain that would be identified as Saccharomyces bayanus by standard DNA barcoding procedures.

  View details for DOI 10.1126/science.aaf3981

  View details for Web of Science ID 000396348900036

  View details for PubMedID 28280149

• A Single-Chain Photoswitchable CRISPR-Cas9 Architecture for Light-Inducible Gene Editing and Transcription A Single-Chain Photoswitchable CRISPR-Cas9 Architecture for Light-Inducible Gene Editing and Transcription Zhou, X. X., Zou, X., Chung, H. K., Gao, Y., Liu, Y., QI, L. S., Lin, M. Z. 2017: 443–48

  Abstract

  Optical control of CRISPR-Cas9-derived proteins would be useful for restricting gene editing or transcriptional regulation to desired times and places. Optical control of Cas9 functions has been achieved with photouncageable unnatural amino acids or by using light-induced protein interactions to reconstitute Cas9-mediated functions from two polypeptides. However, these methods have only been applied to one Cas9 species and have not been used for optical control of different perturbations at two genes. Here, we use photodissociable dimeric fluorescent protein domains to engineer single-chain photoswitchable Cas9 (ps-Cas9) proteins in which the DNA-binding cleft is occluded at baseline and opened upon illumination. This design successfully controlled different species and functional variants of Cas9, mediated transcriptional activation more robustly than previous optogenetic methods, and enabled light-induced transcription of one gene and editing of another in the same cells. Thus, a single-chain photoswitchable architecture provides a general method to control a variety of Cas9-mediated functions.

  View details for DOI 10.1021/acschembio.7b00603

  View details for PubMedCentralID PMC5820652