In this video, we demonstrate the laser-assisted permeation of the protective layer of zona pellucida in mouse fertilized eggs for facilitating lentiviral gene delivery. Lentivirus enables the generation of transgenic animals with a gene of interest stably integrated into their genome.
Protocol
1. Perforation of Mouse Fertilized Eggs with XYClone Laser Setup and calibrate XYClone laser according to the manufacturer's recommendation. Other lasers, typically used for in vitro fertilization, can be substituted for XYClone laser to perforate fertilized eggs. Attach the laser controller box wire to the laser apparatus on the microscope. Attach the laser controller box to the computer running the laser software via a USB port. 3.1.3. Plug in the laser controll…
Representative Results
Figure 1: Laser-Treatment of Mouse Fertilized Eggs. Examples of perforating the zona to produce a hole vs thinning of the zona.
Disclosures
The authors have nothing to disclose.
Materials
CD510B-1 plasmid
System Biosciences
CD510B-1
used to package the lentivirus expressing EF1a-copGFP