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Immunology and Infection
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JoVE Journal Immunology and Infection
Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs
 

Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

Article DOI: 10.3791/55379-v • 11:21 min • September 10th, 2017
September 10th, 2017 •
1, 2, 1, 1, 3, 2, 4, 1, 1
1Department of Pediatrics, Section of Diabetes and Endocrinology, McNair Medical Institute, Baylor College of Medicine, Texas Children's Hospital, 2Benaroya Research Institute at Virginia Mason, 3Center for Human Immunobiology, Baylor College of Medicine, Texas Children's Hospital, 4Department of Pediatrics, Section of Diabetes and Endocrinology, Baylor College of Medicine, Texas Children's Hospital

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Resumen
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The current protocol combines single cell paired human TCR alpha and beta chain sequencing with streamlined generation of retroviral vectors compatible with in vitro and in vivo TCR expression.

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Single Cell TCR Isolation Retroviral Vectors In Vitro Expression Of Human TCRs In Vivo Expression Of Human TCRs Immunology TCR Alpha And Beta Repertoire PCR Efficiency TCR Cloning Two-step PCR Multiplex Reaction Nested Reaction Adapter Sequence Restriction Sites Cloning Into Template Vector Chimeric TCR Construct CD3 Signaling Complex Functional Studies Mirroring Cell Lines Humanized Mouse Models
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