Jul 03, 2023
MED-JET H4 MULTIJET (MJH4M) Transfection Protocol
- Nathan Liu1,2,
- Hani Abd-Ul-Salam2,3,4,
- Noemie Joannette-Lafrance1,2,
- Jingjing Li1,2,
- Karim Menassa5,
- Monzur Murshed1,2,3
- 1Faculty of Medicine and Health Sciences, McGill University, Montreal, Quebec, Canada.;
- 2Shriners Hospital for Children, McGill University, Montreal, Quebec, Canada.;
- 3Faculty of Dental Medicine and Oral Health Sciences, McGill University, Montreal, Quebec, Canada.;
- 4College of Dentistry, Gulf Medical University, Ajman, Ajman, United Arab Emirates.;
- 5Medical International Technologies (MIT Canada) Inc., Montreal, Quebec, Canada.
- Nathan Liu: Nathan Liu and Hani Abd-Ul-Salam contributed equally to the paper. Both are first co-authors.;
- Hani Abd-Ul-Salam: Nathan Liu and Hani Abd-Ul-Salam contributed equally to the paper. Both are first co-authors.;
- Monzur Murshed: Principal Investigator and Corresponding Author;
Protocol Citation: Nathan Liu, Hani Abd-Ul-Salam, Noemie Joannette-Lafrance, Jingjing Li, Karim Menassa, Monzur Murshed 2023. MED-JET H4 MULTIJET (MJH4M) Transfection Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx3ekog8j/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 27, 2023
Last Modified: July 03, 2023
Protocol Integer ID: 84108
Abstract
Transfection, a non-viral method of nucleic acid delivery, often shows poor efficiency in vivo. The needle-based in vivo delivery of transfection reagents can be invasive. Here, we report a non-invasive in vivo gene delivery protocol via the needle-free MED-JET H4 MULTIJET (MJH4M) devices using glucose-based and commercial transfection reagents. The objective of this study is to compare the relative transfection efficiencies of the needle-free system to that of the needle-based delivery method. The highest transfection efficiency was noted using 5% glucose as a delivery agent; a 15-fold increase was observed using MJH4M compared to delivery with needles.
Prepare transfection reagent by diluting renilla-luciferase plasmid DNA (pR-Luc) in 5% glucose (w/v) to a final concentration of 0.1 μg/μl.
Insert the transfection reagent cartridge into the MED-JET H4 MULTIJET (MJH4M) device.
MJH4M device parameters were set to 80 psi and 75 µl.
Adult wild-type C57BL/6 mice were anesthetized via a mixture of inhaled isoflurane and oxygen gas.
The dorsal and ventral regions of the mice hind legs were shaved to expose the skin covering the region below the knee.
The injection landmark was identified on the mice hindleg ventral skin 7mm proximal to the heel.
The tip of the device is positioned directly on the injection landmark and the trigger was pressed. The device was held on the hindleg for 3 seconds to ensure retention of the injected liquid.
Tissue collection occurred 48 hours after the injection. The mice were euthanized using inhaled isoflurane and carbon dioxide followed by cervical dislocation.
Mice hindleg muscles were extracted from the bone and flash frozen in liquid nitrogen.
Luciferase assay and signal normalization were performed to determine the transfection efficiency.