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. 2016 Nov 11;26(12):1349–1352. doi: 10.1038/cr.2016.134

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Copyright © 2016 Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences

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NgAgo-based fabp11a gene knockdown causes eye developmental defects in zebrafish. (A) Schematic diagram showing the structure of NgAgo-2nls for in vitro transcription and microinjection. (B) Schematic diagram showing guide DNA targeting sites (indicated by arrowhead) in the exons of fabp11a gene. The targeting sequences are highlighted in purple and red. (C) Microscopy analysis of eyes in control and FW-guide DNA 2 and NgAgo-2nls mRNA coinjected embryos, dorsal view, with the head regions enlarged and shown on the right. Red arrowheads indicate normal paired eyes. Blue arrowhead indicates one single eye in FW-guide 2 and NgAgo-2nls mRNA coinjected embryo. Green arrowhead indicates a single fused large eye. (D) Statistical analysis of phenotype efficiency in control (0%); NgAgo-2nls mRNA only (0%); FW-guide 2 only (0%); FW-guide 2 and NgAgo-2nls mRNA (29.6%); FW-guide 2 mismatch 1 and NgAgo-2nls mRNA (0%); RV-guide 2 and NgAgo-2nls mRNA (15.4%); FW-guide 2, RV-guide 2 and NgAgo-2nls mRNA (39.7%); FW-guide 2, NgAgo-2nls mRNA and fabp11a mRNA injected embryos (4.6%). One-Way ANOVA; ^**** P < 0.0001; ^* P< 0.05. (E) Relative mRNA levels of zebrafish fabp11a in 30 hpf control; NgAgo-2nls mRNA only; FW-guide 2 only; FW-guide 2 and NgAgo-2nls mRNA; FW-guide 2 mismatch 1 and NgAgo-2nls mRNA; RV-guide 2 and NgAgo-2nls mRNA; FW-guide 2, NgAgo-2nls mRNA and fabp11a mRNA; FW-guide 2, RV-guide 2 and NgAgo-2nls mRNA injected embryos. One-Way ANOVA; ^**** P < 0.0001.