E protein binding at the Tcra enhancer promotes Tcra repertoire diversity

The Tcra gene enhancer (Eα) is required for Tcra rearrangment. Our study assesses the role of E protein binding at Eα. Tcra repertoire sequencing was used to determine whether loss of Eα E protein binding sites alters recombination at Tcra. Pre-selection DP thymocytes (Live Lin- CD4+ CD8+ CD3-) were FACS-sorted from three pairs of Eα(+/+) and Eα(ΔE1ΔE3/ΔE1ΔE3) 3-4 week old mice, six samples total. Each sample was obtained from an individual mouse. Sorted cells were resuspended in TRIzol and RNA purified. RNA was processed to assess Tcra repertoire by high-throughput sequencing.