Endogenous Gene Tagging of pfr2 and pfr5 in Trypanosoma cruzi using CRISPR/Cas9
Document Type
Oral Presentation
Campus where you would like to present
Ellensburg
Event Website
https://digitalcommons.cwu.edu/source
Start Date
15-5-2019
End Date
15-5-2019
Abstract
Trypanosoma cruzi, the causative agent of Chagas disease, is a single-celled parasite that contains a unique structure called the paraflagellar rod (PFR). The PFR is a lattice-like structure of cytoskeletal filaments that extends along the length of the flagellum. The exact functions and protein composition of the PFR has yet to be determined, although several major and minor proteins have been identified. The PFR is not only a complex structure shown to be crucial for cell motility, but it is also part of a group of proteins that have shown to protect mice from a lethal dose of T. cruzi following immunization. PFR5 is a proposed component of the PFR, containing a region of homology (PFR domain) which has been found within several well-characterized PFR proteins. Currently, it is unknown if PFR5 is associated with the PFR, with one of the focuses of this study being on its localization. PFR5 localization is being determined using a new gene tagging technique that utilizes CRISPR/Cas9 technology, with PFR2 localization serving as a proof of principle since its localization is well known. This technique allows for the precise insertion of a small hemagglutinin tag at the end of a gene of interest, with subsequent protein product also containing the tag. Localization of the tagged proteins can then be visualized using fluorescent antibodies. Successful utilization of this technique, as well as localization of the PFR5 protein, will help to further research and understanding of this unique structure.
Recommended Citation
Bryant, Naomi, "Endogenous Gene Tagging of pfr2 and pfr5 in Trypanosoma cruzi using CRISPR/Cas9" (2019). Symposium Of University Research and Creative Expression (SOURCE). 47.
https://digitalcommons.cwu.edu/source/2019/Oralpres/47
Department/Program
Biological Sciences
Slides for SOURCE 2019 presentation Bryant
Additional Files
Naomi Bryant - SOURCE 2019.pptx (14499 kB)Slides for SOURCE 2019 presentation Bryant
Endogenous Gene Tagging of pfr2 and pfr5 in Trypanosoma cruzi using CRISPR/Cas9
Ellensburg
Trypanosoma cruzi, the causative agent of Chagas disease, is a single-celled parasite that contains a unique structure called the paraflagellar rod (PFR). The PFR is a lattice-like structure of cytoskeletal filaments that extends along the length of the flagellum. The exact functions and protein composition of the PFR has yet to be determined, although several major and minor proteins have been identified. The PFR is not only a complex structure shown to be crucial for cell motility, but it is also part of a group of proteins that have shown to protect mice from a lethal dose of T. cruzi following immunization. PFR5 is a proposed component of the PFR, containing a region of homology (PFR domain) which has been found within several well-characterized PFR proteins. Currently, it is unknown if PFR5 is associated with the PFR, with one of the focuses of this study being on its localization. PFR5 localization is being determined using a new gene tagging technique that utilizes CRISPR/Cas9 technology, with PFR2 localization serving as a proof of principle since its localization is well known. This technique allows for the precise insertion of a small hemagglutinin tag at the end of a gene of interest, with subsequent protein product also containing the tag. Localization of the tagged proteins can then be visualized using fluorescent antibodies. Successful utilization of this technique, as well as localization of the PFR5 protein, will help to further research and understanding of this unique structure.
https://digitalcommons.cwu.edu/source/2019/Oralpres/47
Faculty Mentor(s)
Gabrielle Stryker