Martel, D., Beneke, T., Gluenz, E. , Späth, G. F. and Rachidi, N. (2017) Characterisation of casein kinase 1.1 in Leishmania donovani using the CRISPR Cas9 toolkit. BioMed Research International, 2017, 4635605. (doi: 10.1155/2017/4635605) (PMID:29333442) (PMCID:PMC5733176)
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Abstract
The recent adaptation of CRISPR Cas9 genome editing to Leishmania spp. has opened a new era in deciphering Leishmania biology. The method was recently improved using a PCR-based CRISPR Cas9 approach, which eliminated the need for cloning. This new approach, which allows high-throughput gene deletion, was successfully validated in L. mexicana and L. major. In this study, we validated the toolkit in Leishmania donovani targeting the flagellar protein PF16, confirming that the tagged protein localizes to the flagellum and that null mutants lose their motility. We then used the technique to characterise CK1.1, a member of the casein kinase 1 family, which is involved in the regulation of many cellular processes. We showed that CK1.1 is a low-abundance protein present in promastigotes and in amastigotes. We demonstrated that CK1.1 is not essential for promastigote and axenic amastigote survival or for axenic amastigote differentiation, although it may have a role during stationary phase. Altogether, our data validate the use of PCR-based CRISPR Cas9 toolkit in L. donovani, which will be crucial for genetic modification of hamster-derived, disease-relevant parasites.
| Item Type: | Articles |
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| Additional Information: | This work was supported by the ANR-13-ISV3-0009. Daniel Martel was supported by the French Government’s Investissements d’Avenir Program, Laboratoire d’Excellence Integrative Biology of Emerging Infectious Diseases (Grant no. ANR-10-LABX-62-IBEID studentship). Eva Gluenz is a Royal Society University Research Fellow and Tom Beneke was supported by an MRC studentship (15/16_MSD_836338). |
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Gluenz, Dr Eva |
| Authors: | Martel, D., Beneke, T., Gluenz, E., Späth, G. F., and Rachidi, N. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity |
| Journal Name: | BioMed Research International |
| Publisher: | Hindawi Publishing Corporation |
| ISSN: | 2314-6133 |
| ISSN (Online): | 2314-6141 |
| Copyright Holders: | Copyright © 2017 Daniel Martel et al. |
| First Published: | First published in BioMed Research International 2017:4635605 |
| Publisher Policy: | Reproduced under a Creative Commons license |
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