Extended Data Fig. 5. Genetic modifications in the Igh-Igκ hybrid line and single Jκ5 allele line. Related to Fig. 4.
a, Diagram of the strategy for various genetic modifications in the Igh-Igκ hybrid v-Abl line. In brief: (i) Diagram of the Igh-Igκ hybrid line. (ii) Diagram of the Igh-Igκ hybrid-Vκ line which was generated from the Igh-Igκ hybrid line by inverting the whole Vκ locus, mutating both RSSs of DQ52, and deleting all upstream Ds, as illustrated in the diagrams just above. (iii) Diagram of the Igh-Igκ hybrid-Vκ-JκRSS line which was generated from the Igh-Igκ hybrid-Vκ line by replacing the JH1-23RSS with a Jκ5-23RSS. (iv) Diagram of the Igh-Igκ hybrid-Vκ-JκRSS-PKO line which was generated from the Igh-Igκ hybrid-Vκ-JκRSS line by deleting the proximal Vκ domain. See Methods for more details. b, Diagram of Igh-Igκ hybrid-Vκ line, as shown in a(ii). c, Absolute level, and d, relative percentage of individual Vκ-to-JH joins in Igh-Igκ hybrid-Vκ line with JH1 bait. The patterns of distal and middle Vκ usage in the Igh-Igκ hybrid-Vκ line (d) and the single Jκ5-single Igh line (Fig. 4e) are similar (Two-sided Pearson’s r = 0.70, P = 2.2e-21). e, Illustration of single Jκ5-single Igh-JHRSS line, in which Jκ5-23RSS was replaced with JH1-23RSS. f, Absolute level, and g, relative percentage of individual Vκ-to-Jκ joins in single Jκ5-single Igh-JHRSS line with Jκ5 bait. Total rearrangement level in f is 100-fold lower than that in Fig. 4e (P = 0.0006; unpaired, two-sided Welch t-test). Vκ usage data are presented as mean ± s.e.m. from 3 biological repeats.