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. 2019 Apr 2;18:63. doi: 10.1186/s12934-019-1112-2

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© The Author(s) 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 3 — Optimizing strategies of CRISPR–Cas9 system in genome editing of non-conventional yeasts. a The Ku70/80 heterodimer is regulatory DNA-binding subunits of DNA-dependent protein kinase (DNA-PK), which is the main component of the NHEJ system in eukaryotes. Knockout the Ku70 and Ku80 genes can repress the NHEJ system. b Synthetic promoters are generated by placing the pol III promoter sequences immediately upstream of the tRNA. c rDNA tandem array can serve as target loci for multi-copy integrations due to its high copy numbers of head-to-tail repeats. d The T7 polymerase system (T7 polymerase and T7 promoter) can express the sgRNA and enable CRISPR-based genome editing in yeasts