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. 2023 Jan;63(1):239-248.
doi: 10.1111/trf.17197. Epub 2022 Nov 27.

The role of RBC antigen transgene integration sites on RBC biology in mice

Arijita Jash  1   2 , Ariel M Hay  1   2 , J Baodan Collins  1   2 , Jinho Heo  1   2 , Chance John Luckey  1 , Krystalyn E Hudson  3 , James C Zimring  1   2
Affiliations

The role of RBC antigen transgene integration sites on RBC biology in mice

Arijita Jash et al. Transfusion. 2023 Jan.

Abstract

Background: Transgenic mice expressing RBC specific antigens are widely used in mechanistic studies of RBC alloimmunization. Existing RBC donor strains have random transgene integration, potentially disrupting host elements that can confound biological interpretation.

Study design and methods: Integration site and genomic alterations were characterized by both targeted locus amplification and congenic backcrossing in the five most commonly used RBC alloantigen donor strains (KEL-K2hi , KEL-K2med , and KEL-K2lo , and KEL-K1). A targeted transgenic approach was developed to allow RBC specific transgene expression from a safe harbor locus (ROSA26). Alloimmune responses were assessed by transfusing alloantigen expressing RBCs into wild-type recipients and measuring alloantibodies by flow cytometry.

Results/findings: Four of the five analyzed strains had at least one gene disrupted by the transgene integration but none of the disrupted genes are known to be involved in RBC biology. The integration of KEL-K2med potentially altered the immunological properties of RBCs, although the biological significance of the observed changes is unclear. The ROSA26 targeted approach resulted in a single copy of the transgene that maintains RBC specific expression without random disruption of genomic elements.

Conclusion: These findings provide a detailed characterization of genomic disruption by transgene integration found in commonly used RBC donor strains that is relevant to numerous previous publications as well as future studies. With the possible exception of KEL-K2med , transgene integration is not predicted to affect RBC biology in existing models, and new models can avoid this concern using the described targeted transgenic approach.

Keywords: RBC Transfusion; alloimmunization; immunology (other than RBC serology).

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References

REFERENCES

    1. Hendrickson JE, Eisenbarth SC, Tormey CA. Red blood cell alloimmunization: new findings at the bench and new recommendations for the bedside. Curr Opin Hematol. 2016;23:543-9.
    1. Hendrickson JE, Tormey CA. Understanding red blood cell alloimmunization triggers. Hematology Am Soc Hematol Educ Program. 2016;2016:446-51.
    1. Auffray I, Marfatia S, de Jong K, Lee G, Huang CH, Paszty C, et al. Glycophorin A dimerization and band 3 interaction during erythroid membrane biogenesis: in vivo studies in human glycophorin A transgenic mice. Blood. 2001;97:2872-8.
    1. Chaudhuri A, Yuen G, Fang F, Storry J. Development of Duffy transgenic mouse: in vivo expression of human Duffy gene with -33T-->C promoter mutation in non-erythroid tissues. Br J Haematol. 2004;127:356-9.
    1. Gibb DR, Liu J, Natarajan P, Santhanakrishnan M, Madrid DJ, Eisenbarth SC, et al. Type I IFN is necessary and sufficient for inflammation-induced red blood cell alloimmunization in mice. J Immunol. 2017;199:1041-50.

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