Figure 8. Two late‐replicating environments are permissive to a shift toward earlier replication after the site‐specific insertion of a large autonomous replicon.

• A, B
  RT profiles of chromosomal alleles following the targeted integration of a β ^A ‐globin + β‐actin construct into two late‐replicating loci, 1 (A) and 2 (B) in two or three clonal cell lines, respectively. Nascent strands (NS) were quantified by real‐time qPCR in four S‐phase fractions. Specific primer pairs determine the RT profile for the modified allele (With), the wt allele (Without), and both alleles (Both). The endogenous β‐globin locus was analyzed as an early‐replicated control. The modified (Mod) and wild‐type (wt) alleles are shown. Difference –ΔL + ΔE values calculated at the target site following transgene integration are indicated (see Appendix Fig S1). Error bars correspond to the standard deviation for qPCR duplicates. The black vertical bars represent the precise insertion position.