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. Author manuscript; available in PMC: 2019 Jul 20.
Published in final edited form as: Circ Res. 2018 May 4;123(3):335–341. doi: 10.1161/CIRCRESAHA.118.313225

Figure 1. CRISPR/Cas9-mediated Tet2 gene disruption confers a competitive advantage to the hematopoietic stem/progenitor cells.

Figure 1

a. Bone marrow lineage-negative cells from wild type mice were transduced with lentivirus particles expressing Cas9/eGFP and delivered to lethally irradiated wild type mice. b. Flow cytometry analysis of HSPC transduction by lentivirus. Cells are defined as LSK cells (lineage−, c-kit+, Sca-1+) and HSC (CD48−, CD150+ in LSK cells). Transduced cells are GFP positive (n=4). c. Flow cytometry analysis of the peripheral blood at 4 and 16 weeks after reconstitution with bone marrow transduced with Tet2-targeted and control (no Tet2 guide RNA) lentivirus vectors. The percentage of GFP+ cells in both experimental groups are shown (n=6 in both Tet2-indel mice and control mice). Statistical analysis was evaluated by two-way repeated measure ANOVA with Sidak’s multiple comparison tests. d. Results of the TA cloning procedure showing that GFP+ peripheral white blood cells harbor edited Tet2 genes. The wild-type Tet2 sequence is shown for reference. *p<0.05, **p<0.01, ****p<0.0001.