Simple Gene Gun

Figure 1. The blue box contain 13 mm in diameter disk filters placed atop of the pre-filter shown above in the lower left corner of the close up of a Swinnex filter. (Note: the disk filter 3 microns of material passing thru.)

I worked on putting a gene gun together this week with a 16-piece compressor set from Home Depot and a used foot pump. The development of this very simple and inexpensive gene gun seems it might work. On the other hand, I may have come across some problematic obstacles heading down the road of this future apparatus. For instance, the Swinnex chamber with the pre-filter in it has an inlet capacity of 50psi or 3.5bar. The maximum pressure I will need for the successful acceleration of the tungsten particle coated with the desired genes to shoot thru this filter is about 80psi or 5.5bar. Figure 1 above show the device separated into its individual pieces. The red arrow indicates the flow of the air being compressed. I, also, added a pic of the Swinnex filter in the lower right corner of Figure 1.

The attaching of the coupler and other threaded areas needed Teflon tape for proper seal. I tried to use rubber tape that was lying around in the robotics lab room.

All This Reading for Nothing

Diagram1. This poor diagram shows the mechanism of how helium used an force to push the particles of gold coated with the desired DNA in the syringe filter shoots into target of plant tissue. The purple oval shaped circles above the box is where the solenoid would go and the other two is the pressure gauges within the  box.

I started reading an article I found about developing a simple gene gun apparatuses. It included welding pieces of 0.5 centimeters (cm) thick stainless steel sheets into a cube like shape. The open side was covered with 1.5cm thick Plexiglas with 1.0cm thick rubber gasket around the edges. The thick Plexiglas would serve as a door to the cube. Thus, the overall specifications would have a solenoid attached to the top which it served as the mechanism of the gun. Diagram 1 above shows the specifications of the simple gene gun. The vacuum served to silent the nose of the compressed helium shooting thru the shaft of the micro tube of the solenoid. The coated gold particles with the desired genes are shot at a very high acceleration to intervene with the nucleus of the plant cell without disrupting its normal function. In conclusion, the desired DNA is inserted into the DNA of the plant DNA if it was cut in some random area. This will overall make the DNA code for some kind of gene expression.  

Josh on the other hand had a cheaper and less efficient way of making a gene gun with a super soaker. Same concept but cheaper parts and instead of gold for its particle to be coated it will be the chemical powder tungsten. Also, the gas helium would be switched out for nitrogen and carbon dioxide powder.

I Can See Memory

The process of how memory is formed is visually caught in real time for a couple of seconds. The mouse used as an in vivo experiment for this accomplishment had its mRNA or messenger ribonucleic acid tagged with green fluorescent chemicals. Messenger RNA encodes for the beta protein that shapes and structures the brain cell for memory storage. The part of the brain known as the hippocampus was stimulated to watch the mRNA go into process. The hippocampus role is separating events and experience of short term memory to long term memory. Thus, driving a car for the first time creates paths by the mRNA by encoding the nucleus of the neurons to create molecules that shape the path of the dendrites synaptic communication between other neurons.

Figure 1. The process of how the mouse mRNA was tagged with fluorescent green (A). The molecules forming for roles of creating memory of the neuron out to the dendrites (B, C).

Park et al. (2014). Visualization of dynamics of single endogenous mRNA labeled in live mouse. Science. 343 (6169): 422-424. doi: 10.1126/science.1239200