Article Title: Non-photopic and photopic visual cycles differentially regulate immediate, early and late-phases of cone photoreceptor-mediated vision
Figure Lengend Snippet: RPE-expressed rlbp1b is not required cone photopic vision in zebrafish at 5 dpf. A) CRISPR/Cas9 knockout strategy of RPE-expressed rlbp1b . Guides were targeted to exon 2 and exon 3 to induce a 1440 bp deletion in rlbp1b which contains 7 exons. Forward primer 1 (FP1) to reverse primer 1 (RP1) amplifies a 1688 bp wildtype product or 248 bp product when deletion is present. A poison forward primer (FP2) was designed in the middle of the deleted region to amplify a 469 bp wildtype product. B) An agarose gel (1.5%) depicting the presence of the deletion in individual F2 larval genomic DNA. A wildtype control was used to amplify the upper band (469 bp) and gDNA from the injected F0 parent fin was used as a positive control to amplify both upper and lower bands (mosaic). L = 100 bp ladder. C) Gene expression analysis of rlbp1b in rlbp1b −/− eyes. Data were analyzed by unpaired, two-tailed t-tests where **=p
Article Snippet: Genomic DNA extraction and polymerase chain reaction DNA was extracted from injected F0 or germline F2 rlbp1b larvae by boiling the sample at 95°C in NaOH (50 mM) (Sigma Aldrich, UK) before neutralizing with 1/10th Trizma Base (Sigma Aldrich, UK).
Techniques: CRISPR, Knock-Out, Agarose Gel Electrophoresis, Injection, Positive Control, Expressing, Two Tailed Test