Extended Data Fig. 7: Humanized Cbln2 E2 knock-in mouse shows increased Cbln2 expression in neonatal neocortex.
From: Hominini-specific regulation of CBLN2 increases prefrontal spinogenesis
a, Positions of single-guide RNAs (sgRNA1 and 2) to introduce double-strand breaks in the genomic DNA and targeting vector to replace WT mouse Cbln2 E2 region with that of human CBLN2 E2 (humanized hCbln2 E2) are shown. b, Genotyping strategy for F1 mice. Germline transmission in the F1 generation was confirmed by nested PCR using the primer set of mP3/mP4, followed by hP1/P2 as indicated in a. Two founders #13 and #25 were obtained. c, Mice in the following generation were genotyped by PCR with hP1/P2 and mP1/mP2. An example of genotyping for F2 of line #13 mice is shown (c). d, Comparison of Cbln2 expression between WT Cbln2 E2 (Mm;Mm) and hCbln2 E2 (Hs;Hs) neocortex at PD 0 using quantitative reverse transcription-PCR. RNA was extracted from the neocortex following the removal of hippocampus, olfactory bulb and subpallial regions. Two-tailed Student’s t-test; *P = 0.007; Error bars: S.E.M.; N = 5 per genotype. Genotyping were repeated at least two times in b and c.
