A PCR-based method to genotype mice knocked out for all four CD3 subunits, the standard recipient strain for retrogenic TCR/CD3 bone marrow reconstitution technology

Alejandro Ferrer, Adam G Schrum, Diana Gil Pages

Research output: Contribution to journalArticlepeer-review

Abstract

The novel T-cell receptor (TCR)/CD3-retrogenic-reconstitution system represents a very useful strategy for studying TCR/CD3 signaling. Two retroviral vectors containing genes for all six subunits of the TCR/CD3 complex are used to transduce bone marrow precursors and reconstitute lethally irradiated recipient mice. Mice used in this system as bone marrow donors lack all four CD3 subunits (CD3γδÉζ-/-). These mice are generated by crossing the strains CD3ζ-/- and CD3γδÉ-/-, the latter resulting from a knockout construct targeted to CD3É that additionally silences the linked genes, CD3γ and CD3δ. Lacking mature T-cell function, CD3γδ Éζ-/- mice are immunocompromised animals often produced by heterozygous breeding strategies on the C57BL/6 background. As a more rapid and reliable means to identify CD3γδÉζ-/- mice than previously described Northern and Southern blots, we designed polymerase chain reactions to distinguish knockout from wild-type CD3É and CD3ζ alleles, facilitating the identification of CD3γδ Éζ-/- mice.

Original languageEnglish (US)
Pages (from-to)222-226
Number of pages5
JournalBioResearch Open Access
Volume2
Issue number3
DOIs
StatePublished - Jun 1 2013

Keywords

  • DNA; genes; genetic testing; immunology; PCR

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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