Transient Expression of Fluorescent Zebrafish Constructs

To transiently express the following DNA constructs: pT2-huc:Gal4-VP16, pT2-uas:tRFP, uas:memYFP, pT2-uas:SYP-EGFP [60 (link)], and mnx1X3:GAL4, the constructs were diluted to a concentration of 40 ng/μl and microinjected, using a micromanipulator and a PV830 Pneumatic PicoPump (World Precision Instruments, Sarasota, FL), into one-cell-stage eggs. The embryos were kept in Petri dishes, and the pattern of EGFP expression was monitored throughout their development. The fmr1-/- line was kindly provided by Dr. Gordon X. Wang and Prof. Philippe Mourrian (Stanford University, CA). To minimize genetic variations, heterozygous (fmr1+/-) zebrafish were crossed and their progeny genotyped. Either fmr1-/- and its sibling WT adults or their progeny were used in each experiment.

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Shamay-Ramot A., Khermesh K., Porath H.T., Barak M., Pinto Y., Wachtel C., Zilberberg A., Lerer-Goldshtein T., Efroni S., Levanon E.Y, & Appelbaum L. (2015). Fmrp Interacts with Adar and Regulates RNA Editing, Synaptic Density and Locomotor Activity in Zebrafish. PLoS Genetics, 11(12), e1005702.

Publication 2015

PV830 Pneumatic PicoPump

Adults Cell Dishes Dna constructs Eggs Embryos Gal4 vp16 Genetic variations Heterozygous Zebrafish

Corresponding Organization : Bar-Ilan University

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Variable analysis

independent variables

Presence of DNA constructs: pT2-huc:Gal4-VP16, pT2-uas:tRFP, uas:memYFP, pT2-uas:SYP-EGFP [60], and mnx1X3:GAL4

dependent variables

Pattern of EGFP expression throughout zebrafish development

control variables

Concentration of DNA constructs (40 ng/μl)
Microinjection into one-cell-stage eggs using a micromanipulator and a PV830 Pneumatic PicoPump
Zebrafish genotype (fmr1-/- and its sibling WT adults or their progeny)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Genetic control: fmr1-/- and its sibling WT adults or their progeny

Annotations

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