E protein binding at the Tcra enhancer promotes Tcra repertoire diversity

The Tcra gene enhancer (Ea) is required for Tcra rearrangment. Our study assesses the role of E protein binding at Ea. Tcra repertoire sequencing was used to determine whether loss of Ea E protein binding sites alters recombination at Tcra. Overall design: Pre-selection DP thymocytes (Live Lin- CD4+ CD8+ CD3-) were FACS-sorted from three pairs of Ea(+/+) and Ea(?E1?E3/?E1?E3) 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.