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Porous protein microcrystals as a scaffold for nucleic acids and proteins




Masri, Mahmoud, author
Snow, Christopher, advisor
Peebles, Christie, committee member
Takamitsu, Kato, committee member

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Oral delivery of nucleic acids is restricted by a number of limiting factors, particularly protection of guest DNA and RNA from degradation and hydrolysis within the gastrointestinal tract following ingestion. Highly ordered, self-assembling porous protein crystals have been previously explored for enzyme immobilization, and may offer similar advantages for protection and targeted delivery of therapeutic molecules to cells. We have developed a reproducible method for generating sub-micrometer porous microcrystals from CJ, a putative isoprenoid-binding protein from Campylobacter jejuni, which are non-cytotoxic and capable of passively retaining plasmid DNA and small interfering RNA. Furthermore, we have demonstrated that CJ microcrystals are able to deliver functional plasmid and transfect cells in vitro. In addition to nucleic acids, CJ microcrystals are also capable of adsorbing functional Nanoluciferase, and display chemiluminescent activity following exposure to substrate. The results of this study demonstrate that porous protein microcrystals can serve as a suitable scaffold for RNA, DNA, and functional enzymes, and may represent a viable alternative to spherical nanoparticles and liposomes for therapeutic delivery.


2022 Summer.
Includes bibliographical references.

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Embargo Expires: 08/22/2024



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