Fig. 6: Serial, biallelic humanization of TMPRSS2 in ACE2 mouse ES cells.
From: Mouse genome rewriting and tailoring of three important disease loci
a, Schematic of TMPRSS2 humanization design. Top, Tmprss2 gene locus; grey box highlights the region replaced by human TMPRSS2. Bottom, human TMPRSS2 locus; shading highlights the humanization region. The 3′ end of MX1 was defined as the left boundary; for the right boundary, sufficient TMPRSS2 upstream genomic sequence was used to include a putative enhancer. b, Schematic workflow for TMPRSS2 biallelic humanization in ACE2 mouse ES cells. c, Success rate of biallelic humanization in three MC1 mouse ES cell founder lines determined by genotyping PCR: cWZ405 (n = 76), cWZ410 (n = 81) and cWZ412 (n = 13). d, TMPRSS2 copy number determination by qPCR. Copy number determined as in Fig. 3e. WT, wild type. e, Sequencing coverage of TMPRSS2 mouse ES cell clones. Reads mapped to hg38 (top) and mm10 (bottom). f, A double-humanized GREAT-GEMM with both ACE2 and TMPRSS2 was established via tetraploid complementation. Sex: male, two bands (X and Y); female, one band (X only). g, Top, TMPRSS2 expression pattern in double-humanized ACE2 and TMPRSS2 mouse. Bottom, mouse Tmprss2 expression pattern in ACE2-only humanized mouse. Data are mean ± s.d. of three technical replicates.
