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RNA Sequencing of WT, RAG2-KO, and RAG2-KO TCRbeta-Transduced CD4+CD8+ T-Cells Derived From Human Embryonic Stem Cells

ABSTRACT: T-cell development is predicated on the successful rearrangement of the T-cell receptor (TCR) gene loci, which encode for antigen-specific receptors that enable an immune response. Recombination Activating Gene (RAG) 2 is required for TCR gene rearrangements, which occur during specific stages of T-cell development. Here, we differentiated human pluripotent stem cells with a CRISPR/Cas9-directed deletion of the RAG2 gene (RAG2-KO) to elucidate the requirement for the TCRb chain in mediating b-selection during human T-cell development in vitro. In stark contrast to what is seen in mice, human RAG2-KO T-lineage progenitors progressed to the CD4+CD8+ double-positive (DP) stage in the absence of TCRb rearrangements. Nevertheless, RAG2-KO DPs retrovirally-transduced to ectopically express a rearranged TCRb chain showed increased survival and proliferation as compared to control-transduced RAG2-KO DPs. Furthermore, transcriptomic analysis showed that TCRb-transduced and control-transduced RAG2-KO DPs differed in gene pathways related to survival and proliferation. Our results provide new insights as to the distinct requirement for the TCRb chain during human T-cell development.

ORGANISM(S): Homo sapiens

PROVIDER: GSE164276 | GEO | 2021/03/18

REPOSITORIES: GEO

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RNA Sequencing of WT, RAG2-KO, and RAG2-KO TCRbeta-Transduced CD4+CD8+ T-Cells Derived From Human Embryonic Stem Cells

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