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Publication Date

Fall 2017


Niemann Pick Disease Type C1 (NPC1) is a rare lysosomal storage disorder

that affects 1:150,000 people. The disease is characterized by cholesterol

accumulation within lysosomes, as well as clinical cognitive decline and

neurodegeneration. These symptoms can be attributed to a mutation in the

NPC1 gene that leads to the interruption of the intracellular cholesterol

transport. Using CRISPR Cas9 technology, the ability to make a cellular

model with inducible CRISPR gene regulation can give researchers

considerable insight into the cellular pathology of Niemann Pick Disease

Type C1 as well as the ability to apply targeted drug therapy and potentially

lead to drug discovery. We hypothesize that by using bioinformatics, a

CRISPR cas9 system, and genome editing we will make a functioning

HEK293 NPC1 cell model with inducible CRISPR gene regulation. To build

this system, we 1) used online bioinformatics tools to build primers, 2) Hifi

Assembly of mCherry coding sequence with pUC19 plasmid, 3) utilized

gene editing technology with the purpose of inserting the mCherry (a

fluorescent protein) coding DNA sequence into the NPC1 gene and 4)

inserted the built sequence into HEK 293 cells via CRISPR transfection.

Preliminary results show a successful build of the DNA sequence insert

containing the mCherry coding sequence which has been confirmed by

restriction digest and gel electrophoresis. Once completed the model will be

tested in the hopes that it will follow the cellular mechanisms that lead to the

NPC1 cholesterol transporter dysfunction. With the cellular model

successfully made, and functionality proven, future work will include testing

the cells for targeted drug therapy.

Louis Stokes Alliance for Minority Participation (LSAMP) Scholarship



LSAMP Travel Award, ABRCMS Travel Award, Carol J. Major Legacy Poster Award