All animal experiments were performed in accordance with the institutional guidelines.
P0-P1 Actin>GFP1-10 mouse pups were anesthetized on ice until no reflexes were observed. 2 μg / μL NS4B-PQR-GFP11 DNA (from Asian ZIKV strain 150989 or African ZIKV strain Ar41524) and tdTomato DNA dissolved in water was mixed with 0.1% FastGreen dye and loaded into a 1.5 / 0.786 mm (O.D. / I.D.) pulled and beveled
borosilicate glass micropipette (Sutter Instruments). The micropipette was inserted into the right lateral ventricle or ~1.5 mm into the right cortical hemisphere, ~ 1.5 mm laterally from bregma (Figure 4e). Around 200 nL of DNA and dye solution was injected and allowed to diffuse for three minutes.
Platinum tweezer electrodes (Nepagene) were then placed around the pup head such that the negative electrode contacted the injected side of the head. A drop of PBS was used under the electrodes to transmit the pulses. Two sets of nine pulses (100 V, 50 ms, 950 ms apart) separated by three seconds were delivered, and the animal was allowed to recover on a 37°C heated blanket. The pups were returned to their mother once awake and active.
Kays I., & Chen B.E. (2021). Tracking and Measuring Local Protein Synthesis In Vivo.
