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Fig. 2 Generation of a stably-integrated, ubiquitously-expressed QFGal4 driver line. (A) Epifluorescence micrographs showing GFPNLS expression from a Tg(QUAS5x:GFPNLS)hsc143 reporter line crossed to either Tg(ubi:QF2) (left column), Tg(ubi:QF2w) (middle column), or Tg(ubi:QFGal4) (right column) transgenic F0 founder fish. A magnified view of the heart chamber is shown to the right of each merged panel. Tg(ubi:QF2) and Tg(ubi:QF2w) transgenic lines exhibited the strongest activation of the GFPNLS reporter, but also had developmental defects, including heart edema (arrow and dashed line outlining edema in insets). Notably, defects caused by QF2 and QF2w expression were also observed in the absence of the Tg(QUAS5x:GFPNLS)hsc143 transgene (not shown). Most Tg(ubi:QF2w) F1 larvae exhibited strong GFPNLS expression, except progeny from Tg(ubi:QF2w)hsc124, which showed weak variegated GFPNLS expression when crossed a Tg(QUAS5x:GFPNLS)hsc143 reporter line. Tg(ubi:QFGal4) lines showed robust and ubiquitous activation of a GFPNLS reporter line, with no observable toxic effects during larval development. Note that each F1 larvae shown is derived from an independent founder. All larvae shown were imaged at 3 dpf. (B) Epifluorescence micrographs showing GFPNLS expression from a Tg(QUAS5x:GFPNLS)hsc143 reporter line crossed to first generation Tg(ubi:QF2) (left column), Tg(ubi:QF2w) (middle column) or Tg(ubi:QFGal4) transgenic fish (right column). A magnified view of the heart chamber is shown to the right of each merged panel. Tg(ubi:QF2)hsc118 and Tg(ubi:QF2w)hsc124 F2 larvae exhibited weak, mosaic GFPNLS expression (see insets), and Tg(ubi:QF2w)hsc121 exhibited no detectable GFPNLS expression when crossed to a Tg(QUAS5x:GFPNLS)hsc143 reporter line. In contrast, Tg(ubi:QFGal4) F2 larvae exhibited strong, ubiquitous GFPNLS expression. No heart edema was evident in F2 larvae for any of the transgenic lines (see inset panels). Each larvae shown is an F2 established from an independent founder, and all larvae were imaged at 3 dpf. (C) Epifluorescence micrographs showing GFPNLS expression from a Tg(QUAS5x:GFPNLS)hsc143 reporter line crossed to a muscle-specific driver line Tg(acta1:QFGal4)hsc148. Image shows a 3 dpf larvae.
Reprinted from Developmental Biology, 465(2), Burgess, J., Burrows, J.T., Sadhak, R., Chiang, S., Weiss, A., D'Amata, C., Molinaro, A.M., Zhu, S., Long, M., Hu, C., Krause, H.M., Pearson, B.J., An optimized QF-binary expression system for use in zebrafish, 144-156, Copyright (2020) with permission from Elsevier. Full text @ Dev. Biol.