Wang F et al. (2015), Targeted gene disruption in Xenopus laevis usin...

XB-ART-50579

Cell Biosci 2015 Jan 01;5:15. doi: 10.1186/s13578-015-0006-1.

Show Gene links Show Anatomy links

Targeted gene disruption in Xenopus laevis using CRISPR/Cas9.

Wang F , Shi Z , Cui Y , Guo X , Chen Y .

???displayArticle.abstract???

???displayArticle.pubmedLink??? 25897376
???displayArticle.pmcLink??? PMC4403895
???displayArticle.link??? Cell Biosci

Species referenced: Xenopus laevis
Genes referenced: pdia2 tyr
gRNAs referenced: ptf1a gRNA2 ptf1a gRNA3 tyr gRNA14 tyr gRNA16

Phenotypes: Xla Wt + ptf1a gRNA2/3 CRISPR (Fig.3 KN) [+]

???attribute.lit??? ???displayArticles.show???

	Figure 1. pdip â€‰expression is undetectable inâ€‰X. tropicalisÂ ptf1a/p48-null mutant tadpoles.â€‰Whole-mount in situ hybridization analysis ofâ€‰pdipâ€‰expression inÂ F2 X. tropicalisÂ ptf1a/p48â€‰mutant tadpoles (stage 43).â€‰Ventral-lateral view, head to the left. (A, C) About 22% homozygous tadpoles showed complete absence ofâ€‰pdip â€‰expression. (B, D) The remainder uniformly showed strongâ€‰ pdip expression, suggesting normal pancreas development inâ€‰ ptf1a/p48â€‰heterozygous tadpoles.
	Figure 2. CRISPR/Cas9 is effective in targetingâ€‰X. laevisÂ ptf1a/p48.â€‰DNA sequencing data reveal the indel-inducing efficiencies ofâ€‰ptf1a/p48-T1 andâ€‰ptf1a/p48-T2 inâ€‰X. laevisâ€‰embryos. For all panels, the wild-type sequence is shown at the top with the target site highlighted in yellow and the PAM sequence in blue. Red dashes indicate deletions (âˆ’) and lowercase letters in red indicate insertions (+).
	Figure 3. ptf1a/p48-targeted X. laevis G0 tadpoles showed reduction of pdip expression.â€‰Whole-mount in situ hybridization analyses ofâ€‰pdipâ€‰expression (stage 42). Ventral-lateral view, head to the left. (A, B) Wild typeâ€‰X. laevisâ€‰tadpoles. (C, E) Severe reduction. (D, F) Mild reduction. (G, I) Severe reduction. (H, J) Mild reduction. (K, N) Almost complete absence. (L, O) Severe reduction. (M, P) Mild reduction.
	Figure 4. CRISPR/Cas9 is effective in targetingâX. laevisÂ tyrosinase.âDNA sequencing data reveal the indel-inducing efficiencies ofâtyra-T andâtyrb-T inâX. laevisâembryos. For all panels, the wild-type sequence is shown at the top with the target site highlighted in yellow and the PAM sequence in blue. Red dashes indicate deletions (â) and lowercase letters in red indicate insertions (+). The left two panels show data fromâtyra-T andâtyrb-T co-injection and right two panels indicate individual injection results.
	Figure 5. Simultaneous disruption of twoâ€‰ X. laevisÂ tyrosinase â€‰homeologs leads to albinismÂ in G0 tadpoles (stage 42).â€‰Lateral view, head to the left. (A, D) Almost full albinism. (B, E) Severe inhibition of pigmentation. (C, F) Almost normal. (G, J) Wild type. (H, I, K, L) Almost normal.
	pdia2 (protein disulfide isomerase family A member 2 ) gene expression in X. laevis embryos, assayed via in situ hybridization, NF stage 43, lateral view, anterior left, dorsal up.

References [+] :

Afelik, Combined ectopic expression of Pdx1 and Ptf1a/p48 results in the stable conversion of posterior endoderm into endocrine and exocrine pancreatic tissue. 2006, Pubmed, Xenbase