Optimizing CRISPR interference for Conditional Gene Regulation in malaria Parasites

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Date
2019-01-01
Authors
Meis, Ellen
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Honors Projects and Posters
University Honors Program

The Honors project is potentially the most valuable component of an Honors education. Typically Honors students choose to do their projects in their area of study, but some will pick a topic of interest unrelated to their major.

The Honors Program requires that the project be presented at a poster presentation event. Poster presentations are held each semester. Most students present during their senior year, but may do so earlier if their honors project has been completed.

This site presents project descriptions and selected posters for Honors projects completed since the Fall 2015 semester.

Department
Ecology, Evolutionary, and Organismal Biology
Abstract

To continue fighting malaria, identification of new drug targets or life cycle intervention points and a better understanding of malaria parasite biology are crucial. While the ability to genetically manipulate the parasite in order to assess gene function is key to this goal, the tools to study important parasite genes are limited. CRISPR interference (CRISPRi) has emerged as a powerful and simple approach for conditional gene knockdown using enzymatically inactive Cas9 (dCas9). This project aims to develop and optimize a CRISPRi system in the most virulent human malaria parasite, Plasmodium falciparum, by determining optimal guide RNA design to achieve maximum target gene knockdown. To this end, we established a dual luciferase reporter system to test the activity of a tiled array of guide RNAs around the start codon of a gene target. We hypothesize that as the guide RNA location approaches the start codon, knockdown via dCas9 interference with transcription by RNA polymerase will increase. After determining the optimal guide RNA placement, the system can be used to screen putative transcriptional effector proteins to enhance knockdown. This system has the potential to provide a robust approach for studying essential P. falciparum genes with increased throughput. Current results will be discussed.

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