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Fig. 3 mNG211 tagging by CRISPR/Cas-directed gene editing. A. Schematic of CRISPR/Cas-directed mNG211 insertion into target genes. Purple, endogenous exon sequence. Green, mNG211. Yellow, linker (LK). ATG, start codon. Arrows denote primers used in B. B. mNG211 insertion was assessed by PCR. The primers used correspond to the arrows shown in A. bp, base pairs. C. Amino acid sequences of wild-type, predicted mNG211 fusions, and recovered alleles for Tubb4b and Krt8. Mismatches between the predicted and recovered sequences are highlighted in red. Asterisks, stop codons. D?I. Representative images of mNG211-tubb4b (D?F) and krt8-mNG211 (G?I) embryos injected with mNG21-10 mRNA (D?E, G?H) or uninjected (F, I). Maximum projections of confocal z-stacks. Images were acquired at 24 h post-fertilization. Images in F and I have been overexposed to emphasize lack of fluorescence. Scale bars, 50 ?m.
Reprinted from Developmental Biology, 514, Ligunas, G.D., Paniagua, G., LaBelle, J., Ramos-Martinez, A., Shen, K., Gerlt, E.H., Aguilar, K., Nguyen, N., Materna, S.C., Woo, S., Tissue-specific and endogenous protein labeling with split fluorescent proteins, 109-116, Copyright (2024) with permission from Elsevier. Full text @ Dev. Biol.