A new technique, called nanoinjection, is being developed to insert foreign DNA into a living cell. Such DNA transfection is commonly used to create transgenic organisms vital to the study of genetics, immunology, and many other biological sciences. In nanoinjection, DNA, which has a net negative charge, is electrically attracted to a micromachined lance. The lance then pierces the cell membranes, and the voltage on the lance is reversed, repelling the DNA into the cell. It is shown that DNA motion is strongly correlated to ion transport through a process called electrophoresis. Gel electrophoresis is used to move DNA using an electric field through a gel matrix (electrolytic solution). Understanding and using electrophoretic principals, a mathematical model was created to predict the motion (trajectory) of DNA particles as they are attracted to and repulsed from the nanoinjector lance. This work describes the protocol and presents the results for DNA motion experiments using fabricated gel electrophoresis devices. Electrophoretic systems commonly use metal electrodes in their construction. This work explores and reports the differences in electrophoretic motion of DNA (decomposition voltage, electrical field, etc.) when one electrode is constructed from a semiconductor, silicon rather than metal. Experimental results are used to update and validate the mathematical model to reflect the differences in material selection. Accurately predicting DNA motion is crucial for nanoinjection. The mathematical model allows investigation of the attraction/repulsion process by varying specific parameters. Result show that the ground electrode placement, lance orientation and lance penetration significantly affect attraction or repulsion efficiency while the gap, lance direction, lance tip width, lance tip half angle and lance tip height do not. It is also shown that the electric field around the lance is sufficient to cause localized electroporation of cell membranes, which may significantly improve the efficiency of transport.

A deeper understanding of human development and disease is made possible partly through the study of genetically modified model organisms, such as the common mouse (Mus musculus). By genetically modifying such model organisms, scientists can activate, deactivate, or highlight particular characteristics. A genetically modified animal is generated by adding exogenous (foreign) genetic material to one or more embryonic cells at their earliest stages of development. Frequently, this exogenous genetic material consists of specially engineered DNA, which is introduced into a fertilized egg cell (zygote). When successfully introduced into the zygote, the exogenous DNA will be incorporated into the cell's own genome, and the animal that develops from the zygote will exhibit the genetic modification in all of its cells. The current devices and methods for generating genetically modified animals are inefficient, and/or difficult to use. The most common and efficient method for inserting new DNA into zygotes is by directly injecting a DNA solution through a tiny glass tube into the cell in a process called microinjection. Unfortunately, microinjection is quite inefficient (success rates are commonly between 1 and 5%), but often it is the only method for inserting DNA into eggs, zygotes, or early stage embryos. This thesis presents the design and testing of a micrometer sale, pumpless microelectromechanical system (MEMS) nanoinjector. Rather than use pumps and capillaries, the nanoinjector employs electrostatic charges to attract and repel DNA onto and off of the surface of a solid lance. The nanoinjector also includes a mechanical system for constraining the target cells during injection. Initial testing indicates the nanoinjector does not decrease cell viability, and it has a very high initial success rate (up to 90%). With the addition of an on-chip actuator, the nanoinjector could be packaged as an inexpensive, fully automated system, enabling efficient, high volume genetic modification of developing animals. Such a device would greatly increase the ease and speed of generating the model organisms needed to study such critical diseases such as Alzheimer's disease, cancer, and diabetes.