Project Title

Development and Application of TAT-CaM in Crypthecodinium cohnii

Presenters

Academic department under which the project should be listed

CSM - Molecular and Cellular Biology

Faculty Sponsor Name

Jennifer Cooper

We do not have human subjects.

Abstract (300 words maximum)

Crypthecodinium cohnii is a unique dinoflagellate that is both an important part of the environment, and a valuable source of the omega-3 fatty acid docosahexaenoic acid (DHA). Outside of its use in DHA production, little is known about the organism. We are currently attempting to manipulate and investigate this dinoflagellate by using the Cell-Penetrating Peptide (CPP). The specific CPP we are using is TAT-CaM, which is a hybrid protein made of a TAT, a protein taken from HIV that enters the cell via endocytosis, and calmodulin (CaM) a protein that releases its bound constituents in low calcium environments like the interior of the cell. If this can be done, it will allow deeper study of C. cohnii, such as a further exploration of the species’ unique DHA content, and an investigation of their histone deficient DNA. We have established that C.cohnii is not killed in the process of CPP by using detailed hemocytometer viability counts. We have also found promising signs of successful penetration of the cells using TAT-CaM, which has been bound to Dylight-550 labelled myoglobin. Confocal microscopy results indicate that TAT-CaM is associating with the cell. Future work will focus on optimizing Tat-Cam CPP in C. cohnii.

Project Type

Poster

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Development and Application of TAT-CaM in Crypthecodinium cohnii

Crypthecodinium cohnii is a unique dinoflagellate that is both an important part of the environment, and a valuable source of the omega-3 fatty acid docosahexaenoic acid (DHA). Outside of its use in DHA production, little is known about the organism. We are currently attempting to manipulate and investigate this dinoflagellate by using the Cell-Penetrating Peptide (CPP). The specific CPP we are using is TAT-CaM, which is a hybrid protein made of a TAT, a protein taken from HIV that enters the cell via endocytosis, and calmodulin (CaM) a protein that releases its bound constituents in low calcium environments like the interior of the cell. If this can be done, it will allow deeper study of C. cohnii, such as a further exploration of the species’ unique DHA content, and an investigation of their histone deficient DNA. We have established that C.cohnii is not killed in the process of CPP by using detailed hemocytometer viability counts. We have also found promising signs of successful penetration of the cells using TAT-CaM, which has been bound to Dylight-550 labelled myoglobin. Confocal microscopy results indicate that TAT-CaM is associating with the cell. Future work will focus on optimizing Tat-Cam CPP in C. cohnii.