Generation of Transgenic Caenorhabditis elegans by Aerosol Beam Injection

by Elizabeth Ahrendsen, Shane Clune, Alyssa J. Hudnall, and Alex M. Abel
Faculty advisor: Dr. James Hampton

The purpose of our research project is to effectively transform the nematode C. elegans via Aerosol Beam Injection.  Aerosol Beam Injection may be an alternative to particle bombardment or microinjection as a transformation method.  C. elegans is a useful genetic model organism and a simple and effective way to transform them would benefit many genetic researchers.

Aerosol Beam Injection has been used to transform E. coli, yeast, and plants.  The device aerosolizes DNA and uses a pressure differential system to force the aerosolized mixture toward the organism at approximately the speed of sound.  To test the effectiveness of Aerosol Beam Injection in nematodes, we attempted to resinsert a gene into a knockout strain of C. elegans called unc-119.  The unc-119 gene affects a collagen protein of C. elegans, causing the mutant nematodes to move in an uncoordinated manner and prevents their larvae from entering into a suspended animation state known as dauer.  To rescue the phenotype we inserted a plasmid containing the unc-119 gene.  This plasmid also provided us with an internal control since it will also impart ampicillin resistance to E. coli.

Different parameters were tested; including time each section of the plate was subjected to bombardment and concentration of TRIS buffer used to suspend the DNA.  Other things to be considered are age of the worms being bombarded and care of the worms after we attempted to insert the plasmid.