Team:Tec-Chihuahua
Project Description
Genetic transformation is the introduction of foreign genetic material in prokaryotes (bacteria) and non animal eukaryotic cells such as fungi, algae or plants and transfection when introduced into animal cells. Some techniques used in the laboratory for transforming and transfecting cells are based on the production of competent cells by chemical and physical treatments; however these treatments have low efficiencies. Other methods use bacterial and / or viral vectors capable of infecting the cell of interest and introduce exogenous genetic material. Despite being effective they have some disadvantages as not all species are susceptible to being infected by these microorganisms, and it is not accessible technique for laboratories with little microbiology experience. Techniques using DNA microinjection can be extremely expensive and have very low efficiency. In recent years they have sought new strategies for effective transformation of cells at low cost. One of these strategies is the use of nanotechnology, which has the potential of creating new structures with the ability to cross cell membranes and increase solubility, stability and bioavailability of biomolecules, thereby improving efficiency of release. Carbon nanotubes (CNTs) are cylindrical nanoparticles that are used in the medical field as biological carriers. Functionalization of CNTs with DNA (DNA-CNTs) has been addressed in several research fields for generating gene delivery systems.
The aim of this project is to evaluate the efficiency of DNA-CNTs transformation and transfection in different cell systems. For this reason CNTs will be synthesized with a size of 40-50 nm in diameter and a length of <2 µm. The CNTs will be functionalized with a plasmid DNA, using a polyethylenimine as a bridge between the CNT and DNA, which contains a gene of the green fluorescent protein as a marker. Subsequently the transformation efficiency of Escherichia coli cultures, embryos in early development of Bos Taurus and calluses of Nicotiana tabacum will be evaluated and their efficiencies will be compared with the traditional methods used in the laboratory.