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Description
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is a novel capability inherent in some bacteria that allows for the alteration of DNA. In bacteria, this capability is used to defend against viruses, but in human DNA, CRISPR can be used for a variety of purposes. Paired with a CAS-9 protein, CRISPR can be used to create programmable DNA insertions or deletions. CRISPR CAS-9 will replace existing nonspecific, clumsy DNA-editing mechanisms, and is poised to revolutionize the biological industry. There is debate about the ethical implications of using CRISPR, but there is no doubt about its impending prevalence in disease treatment and personalized therapeutics. The technology is reliant on novel mechanisms to transfect a cell—meaning insert the CRISPR CAS-9 protein into the desired cell— and many independent researchers are exploring mechanisms to improve transfection. Micro Electrical Mechanical Systems (MEMS) provide an improvement to current state of the art. As a basis for rapidly prototyping MEMS devices, SU-8 is a suitable material to create a micro electroporation device for improved transfection for CRISPR applications. SU-8 can enable rapid prototyping early in the design process while providing process pathways to semiconductor material properties. In this thesis, we introduce a fabrication method that utilizes a polymer material—SU-8— that can then serve as the final product: a glassy carbon micro electroporation fluidic chip. The key outcome of this work is the development of new designs and novel fabrication techniques to produce devices of microscale electroporation devices demonstrating electric field strength of 0.3-4.2 KV/M while consisting of multiple layers of the polymer that will become a multilayer glassy carbon structure to improve sample preparation for CRISPR applications.
