Figure 1. TALEN-mediated inactivation of the gene for human Rap1.
(A) Schematic of human Rap1, the TERF2IP locus, the targeting construct, and the resulting knockout allele. F1, R1, and R2neo: PCR primers for genotyping. Arrows in bold: TALEN binding and cut sites.
(B) PCR genotyping of the TERF2IP gene and western blotting for Rap1 in the indicated clones. +, WT allele; Δ, targeted allele; mut, mutation resulting in loss of Rap1.
(C) Southern blot of EcoRI (RI)- or BglII/EcoRV (RV)-digested genomic DNA from targeted HT1080 clones. Probe shown in (A).
(D) Telomeric ChIP of WT and targeted HT1080 clones. Duplicate dot-blots were probed for telomeric or Alu repeats.
(E) Average percentage of telomeric DNA recovered in ChIPs with the indicated antibodies (two independent experiments). Error bars: SEMs.
(F) Schematic illustrating NHEJ and HDR after TALENs cutting.
(G) Table indicating number of Rap1 knockout clones acquired and the genetic alterations in TERF2IP.
See also Figure S1.