Journal: Structure (London, England : 1993)

Article Title: High Resolution Crystal Structure and Chemical Screening Reveal Pantothenate Kinase as a New Target for Antifungal Development

doi: 10.1016/j.str.2022.09.001

Figure Lengend Snippet: A) Schematic representation of the Coenzyme A and ergosterol biosynthesis pathways in fungi. B) AfPanK native promoter (PpanK) replacement strategy with CRISPR-Cas9 system in A. fumigatus locus. Hygromycin resistance is conferred by the hph gene encoding a bacterial hygromycin B phosphotransferase while pNiiA is a nitrogen-regulated A. fumigatus promoter. pNiiA promoter is induced by nitrate or other secondary nitrogen source and repressed by ammonium or other primary nitrogen source. Primers for sequence validation are numbered P1-P3. PCR screen of hygromycin resistant transformants using two pair of primers P1/P2 and P3/P2 is provided in supplementary information. C) Radial growth capture of A. fumigatus wild-type and PniiA-panK strains in solid media under two different nitrogen sources. D) Expression of PanK in A. fumigatus wild-type and PniiA-panK strains under two different nitrogen sources. PanK expression was measured using rtPCR and normalized to basal actin expression. The data represent two biological replicates (n=2) and the plotted graphs represent the average ± SEM. E) Radial growth data of A. fumigatus wild-type and PniiA-panK strains in solid media under two different nitrogen sources. F) Solid medium assay captures of A. fumigatus wild-type and Pnii-ApanK strains in media with increasing concentrations of voriconazole. See also Table S2.

Article Snippet: All CRISPR reagents, including the crRNAs, tracrRNA, and Cas9 nuclease V3 and associated buffers were from Integrated DNA Technologies (IDT).

Techniques: CRISPR, Sequencing, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: eLife

Article Title: Natural variation in C. elegans arsenic toxicity is explained by differences in branched chain amino acid metabolism

doi: 10.7554/eLife.40260

Figure Lengend Snippet:

Article Snippet: Cas9 protein (IDT, 1074182) was injected at 23 µM.

Techniques: Sequencing, Transfection, Construct, Plasmid Preparation