Figure 4: Analysis of 1.6 F1 offspring for BAC transmission and hSIRPA protein expression. | Scientific Reports

Figure 4: Analysis of 1.6 F1 offspring for BAC transmission and hSIRPA protein expression.

From: Comparative Analysis of piggyBac, CRISPR/Cas9 and TALEN Mediated BAC Transgenesis in the Zygote for the Generation of Humanized SIRPA Rats

Figure 4

(A) The PCR analysis shows human SIRPA BAC detection in 9 out of 13 offspring from the founder 1.6. Genomic DNA from founder 5.7 was used as a positive control. (B) Animals 1.6 F1.3 and 1.6 F1.4 are F1 offspring animals representative of PCR SIRPA-BAC positive or negative animals, respectively. Peripheral blood mononuclear cells (PBMCs) were first gated by morphology (dot plot SSC-FSC), then CD11b+ cells (mostly expressed by monocytes) were labelled with a rat anti-CD11b antibody (dot plot CD11b-FSC) and finally, cells were stained with combinations of human and rat anti-SIRPα antibodies (left dot-plots) or human anti-SIRPA antibodies and human CD47-Fc (right dot-plots). The large majority of rat monocytes from BAC-SIRPA-TIR+ transgenic rats expressed human SIRPα, as detected using both anti-human SIRPα antibodies and human CD47-Fc whereas all monocytes from BAC-SIRPA-TIR− transgenic rats were negative for both labels.

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