MOESM1 of Efficient CRISPR–Cas9 mediated multiplex genome editing in yeasts
Creators: name: Wang, Laiyou
name: Deng, Aihua
name: Zhang, Yun
name: Liu, Shuwen
name: Liang, Yong
name: Bai, Hua
name: Cui, Di
name: Qiu, Qidi
name: Shang, Xiuling
name: Yang, Zhao
name: He, Xiuping
name: Wen, Tingyi
Publisher: name: figshare
Year: 2018
Subjects: subject: Biochemistry
subject: Space Science
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Genetics
subject: Molecular Biology
subject: Evolutionary Biology
subject: Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: Cancer
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Virology
subject: Computational Biology

description: Additional file 1: Table S1. Strains and plasmids used in this study. Table S2. Primers used in this study. Table S3. Potential off-target sites of CRISPR–Cas9 mediated point mutation in the gene OpURA3. Table S4. Editing efficiencies mediated by CRISPR–Cas9 in O. polymorpha. Table S5. Editing efficiencies mediated by endogenous HRS in O. polymorpha. Table S6. Editing efficiencies of CRISPR–Cas9-assisted genome engineering methods in different yeasts. Figure S1. Analysis of the editing efficiency mediated by endogenous HRS at the OpADE2 site by cell growth phenotype on YPD and SC without adenine (SC-ADE) plates. Figure S2. PCR identifications of the deletion of OpLEU2 (A) and OpURA3 (B) genes, respectively. Figure S3. Evictions of the linearized gRNA delivery vector and the linearized Cas9 protein expression vector after gene editing. Figure S4. Verification of gene deletions by auxotrophic phenotype analysis. Figure S5. Effect of HA (homologous arm) on editing efficiency of CRISPR–Cas9 mediated gene deletion in O. polymorpha. Figure S6. PCR identifications of simultaneous deletions of genes OpLEU2, OpHIS3 and OpURA3. Figure S7. Verification of multiplex knock-outs by auxotrophic phenotype analysis. Figure S8. The identification of the point mutation by cell growth phenotype. The YPD plate and SC plate without uracil (SC-URA). Figure S9. Verifications of point mutation of the gene OpURA3. Figure S10. DNA sequencing of similar genomic loci of point mutation site in the gene OpURA3 of the mutant OP040 (OP001 OpURA3G73T). Figure S11. PCR identifications of gfpmut3a expression cassette separately integrated at OpLEU2 (A), OpHIS3 (B) and OpURA3 (C) loci. Figure S12. Analysis of editing efficiencies mediated by endogenous homologous recombination system at the gene OpLEU2 (A), OpHIS3 (B) and OpURA3 (C) sites by cell growth phenotype. Figure S13. Editing efficiencies at three gene sites by two different methods. Figure S14. PCR identifications of simultaneously multi-loci genomic integration. Figure S15. PCR identification of gfpmut3a expression cassette by multi-copy integration at rDNA cluster in O. polymorpha. Figure S16. Flow cytometry analysis of the expression of gfpmut3a in eight randomly selected O. polymorpha colonies. Figure S17. Stability of multi-copy integration of gfpmut3a at rDNA repeats in the mutant OP025 upon continuous culture for 96 h. Figure S18. PCR identification of gfpmut3a expression cassette by multi-copy integration at rDNA cluster in S. cerevisiae. Figure S19. Stability of multi-copy integration of gfpmut3a at rDNA repeats in the mutant SC007 upon continuous culture for 96 h.