tet2 mouse model  (New England Biolabs)

Journal: bioRxiv

Article Title: Systematic functional dissection of germline noncoding risk variants impacting clonal hematopoiesis

doi: 10.64898/2026.01.21.700897

Figure Lengend Snippet: (A) Schematic of lentiviral MSI2 overexpression constructs and Western blot confirming MSI2 overexpression in K562 cells. (B) Setup of MSI2 overexpression experiments in murine Tet2 -/- hematopoietic stem and progenitor cells (HSPCs). (C) Colony replating assays of sorted GFP + Lin - Sca-1 + c-Kit + (LSK) cells shows increased self-renewal of Tet2 -/- HSPCs upon MSI2 upregulation. (D) Venn diagram showing common and unique differentially expressed genes across different genotypes identified by RNA-Seq (FDR <0.05 and |log2FC| >1.5). “WT MSI2” indicates wild-type HSPCs transduced with MSI2 overexpressing vector; “Tet2 -/- MSI2” indicates Tet2 -/- HSPCs transduced with MSI2 overexpressing vector; “Tet2 -/- HMD” indicates Tet2 -/- HSPCs transduced with empty HMD vector. All groups are compared against WT HSPCs transduced with empty HMD vector, “WT HMD” (n=2 replicates each group). (E) Heatmap showing relative expression of Wnt polarity genes across all groups. (F) Heatmap showing relative expression of Cell cycle genes across groups. (G) Gene set enrichment analysis of MSI2 HyperTRIBE binding targets in Tet2 -/- MSI2 vs. WT HMD. All data are means ± SDs. *p < 0.05, **p < 0.01.

Article Snippet: Bulk RNA-seq was performed in two biological replicates using primary HSPCs (LSK cells) isolated from a well-established TET2 mouse model. Total RNA was extracted using the RNeasy Mini Kit, and poly(A) enrichment and library preparation were carried out with the NEBNext Ultra II Directional RNA Library Prep Kit.

Techniques: Over Expression, Construct, Western Blot, RNA Sequencing, Transduction, Plasmid Preparation, Expressing, Binding Assay

Journal: bioRxiv

Article Title: Systematic functional dissection of germline noncoding risk variants impacting clonal hematopoiesis

doi: 10.64898/2026.01.21.700897

Figure Lengend Snippet: (A) CFU-C replating assay shows the colonies number from sorted murine GFP+ TET2+/- HSPCs. (B-C) Colony images of 2 nd plating from TET2+/- and TET2-/-HSPCs, respectively. (D) Heatmap showing expression of genes across all genotypes highlighting genes uniquely up- and down-regulated in the KO HMD group (E) Heatmap represents mitochondrial electron transport chain genes expression in different experimental groups. (F) Volcano plots show DEGs expression compared WT MSI2 vs WT HMD, Tet2-/- HMD vs WT HMD, Tet2-/- MSI2 vs Tet2-/- HMD, and Tet2-/- MSI2 vs WT HMD. (G) Gene ontology analysis of downregulated and upregulated pathways between Tet2 -/- MSI2 vs. WT HMD.

Article Snippet: Bulk RNA-seq was performed in two biological replicates using primary HSPCs (LSK cells) isolated from a well-established TET2 mouse model. Total RNA was extracted using the RNeasy Mini Kit, and poly(A) enrichment and library preparation were carried out with the NEBNext Ultra II Directional RNA Library Prep Kit.

Techniques: Expressing

Journal: Nature Communications

Article Title: Vitamin C epigenetically controls osteogenesis and bone mineralization

doi: 10.1038/s41467-022-32915-8

Figure Lengend Snippet: a TET2 western blot after CRISPR-dCas9-mediated TET2 o/e (overexpression) and b 5hmC dot blot under the same conditions. c ECM mineralization after TET2 o/e at day 21 of differentiation. d Osteoblastic gene expression during TET2 o/e. Bar graphs represent mean ± SD; * p < 0.05; ** p < 0.01; *** p < 0.001. Unpaired two-tailed t test in ( c ); two-way ANOVA analysis with Tukey’s multiple comparison tests, groups compared to D0 Ctrl or as marked in d . N = 3 ( c , d ) per group, from biologically independent experiments. Source data as well as exact p = values for all comparisons in d are provided in the Source Data File.

Article Snippet: The Tet2 fl/fl mouse model harbors two loxP sites flanking exon 3 and was purchased from The Jackson Laboratory ( Tet2 tm1.1Iaai ; #017573).

Techniques: Western Blot, CRISPR, Over Expression, Dot Blot, Gene Expression, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: Vitamin C epigenetically controls osteogenesis and bone mineralization

doi: 10.1038/s41467-022-32915-8

Figure Lengend Snippet: a Schematic representation of the mouse models used. Bone tissue was collected at 11 weeks; cKO, conditional knockout. b Prrx1 driven TET1 and TET2 knockdown efficiencies in femurs of Tet1/2 double knockout mice. c 5hmC dot blot and relative quantification. d–f hMeDIP-Seq of femurs from Tet1/2 double knockout mice. d Principal component (PC) analysis and 5hmC peaks around selected bone specific genes ( e ). f Correlation between 5hmC occupancy (TSS + /−30 kb) and gene expression as measured by RNA-Seq; FC, fold change. g Expression of Bone60 genes in conditional Tet1 , Tet2 or Tet1/2 femurs versus Ctrl femurs. h Wordcloud representing phenotypes associated with distal intergenic loci with the strongest 5hmC loss in double knockout vs Ctrl femurs as analyzed by GREAT analysis; FDR, false discovery rate. i µCT analysis of L5 spine and relative quantification; TV total volume, BV bone volume, Tb.N. trabecular number, Tb.Th. trabecular thickness, Tb.Sp. trabecular separation ( j ). Bar graphs represent mean ± SD; * p < 0.05; ** p < 0.01; *** p < 0.001. One-way ANOVA analysis with Tukey’s ( c ) and Dunnett’s ( j ) multiple comparison tests. FDR-adjusted two-tailed, unpaired t tests in g , two-sided Fishers exact test ( f ). In c , Ctrl vs Tet1/2 cKO *** p = 0.0009, Tet1 cKO vs Tet1/2 cKO *** p = < 0.0001, Tet1 cKO vs Tet2 cKO ** p = 0.005, Tet2 cKO vs Tet1/2 cKO * p = 0.0118. N = 5 ( c ) & n = 6 per group in j from biologically indepe n dent animals. Source data are provided as a Source Data File.

Article Snippet: The Tet2 fl/fl mouse model harbors two loxP sites flanking exon 3 and was purchased from The Jackson Laboratory ( Tet2 tm1.1Iaai ; #017573).

Techniques: Knock-Out, Knockdown, Double Knockout, Dot Blot, Quantitative Proteomics, Gene Expression, RNA Sequencing, Expressing, Comparison, Two Tailed Test

Journal: Cancers

Article Title: E-Cigarette Exposure Decreases Bone Marrow Hematopoietic Progenitor Cells

doi: 10.3390/cancers12082292

Figure Lengend Snippet: E-cigarette exposure increases peripheral blood chimerism of JAK2 V617F mutant cells. ( A ) Experimental design. Relative change in peripheral blood chimerism of ( B ) JAK2 V617F mutant and ( C ) TET2 −/− cells from pre-exposure. ( D ) Relative contribution of JAK2 V617F to HSPC populations after 2 months of exposure. Data are shown as mean ± SEM. n = 5–10 mice/group.

Article Snippet: The TET2 knockout ( TET2 −/− ) mouse model has been described [ ] and heterozygous breeding pair was purchased from The Jackson Laboratory (JAX stock #023359).

Techniques: Mutagenesis