Journal: bioRxiv
Article Title: Human-chimpanzee tetraploid system defines mechanisms of species-specific neural gene regulation
doi: 10.1101/2025.03.31.646367
Figure Lengend Snippet: (A) Genomic interval containing TNIK and RE- TNIK in hg38 coordinates. (B) RE- TNIK is a cis -regulated open chromatin region with increased chromatin accessibility in chimpanzee compared to human NPCs. The location of the three gRNAs used to target RE- TNIK are indicated. (C) 50µM of KY-05009, a TNIK inhibitor, decreased luciferase activity from the Wnt reporter TOPflash in the presence of 50µM of CHIR-99021, a Wnt agonist (red). KY-05009 had no effect on luciferase activity from the control reporter FOPflash (gray). (D) Volcano plot of differential gene expression between chimpanzee NPCs infected with gRNAs targeting RE- TNIK and NTC gRNAs. Genes significant at 5% FDR are in red. Highly significant genes, as well as examples of genes associated with Wnt signaling, are labelled. (E) Significant terms from gene-set enrichment analysis of differential gene expression between chimpanzee NPCs infected with gRNAs targeting RE- TNIK and NTC gRNAs. See also Fig. S4, Table S4, and Materials and Methods.
Article Snippet: For the Wnt reporter assays, NPCs were transfected with 0.2µl Lipofectamine Stem Transfection Reagent, 2ng of pNL1.1.TK (Promega, cat #N1501), and 198ng of TOPflash (Addgene, cat #12456) or FOPflash (Addgene, cat #12457) ( ) in 10µl Opti-MEM.
Techniques: Luciferase, Activity Assay, Control, Gene Expression, Infection