ARMIYAW SEBASTIAN NASAMU, Ph.D

SUMMARY

• More than a decade of experience in molecular biology, microbiology, cell biology, genome editing, and drug discovery.
• Genetically engineered Plasmodium and Toxoplasma parasites for conditional gene regulation (knockouts/knock-ins), mutagenesis, and epitope tagging using CRISPR/Cas9 technology.
• Pioneered the use of linear plasmids for facile cloning of AT-rich DNA in Plasmodium falciparum.
• Combined linear plasmid technology with CRISPR/Cas9 and conditional knockdown technology to regulate gene expression in Plasmodium falciparum parasites.
• Leveraged these genetic tools to study two enigmatic proteases (Plasmepsin IX and X) in Plasmodium falciparum.
• Characterized protease expression, localization, function, and substrates, and identified protease inhibitors now in clinical trials.

EDUCATION

• PhD in Microbiology and Microbial Pathogenesis – Dan Goldberg Lab (2021)
  Washington University School of Medicine in St. Louis, St. Louis, MO
• MS in Toxicology – Jacquin Niles Lab (2015)
  Massachusetts Institute of Technology, Cambridge, MA
• BS in Biology and Chemistry (2013)
  Massachusetts Institute of Technology, Cambridge, MA

WORK EXPERIENCE

Postdoctoral Associate – Kevin Brown Lab 7/2023 – 7/2024
Oklahoma University Health Science Center

• Used CRISPR/RNP technology to perform complementation of an essential gene in Toxoplasma gondii.
• Demonstrated that a specific amino acid is crucial for the function of this essential gene. Generated mutant plasmids to interrogate important domains of the gene.

Postdoctoral Associate – Stefan Kappe Lab 9/2021 – 5/2023
Seattle Children's Hospital

• Leveraged expertise in genetic engineering to create specific mutant Plasmodium falciparum parasites.
• Used these parasites to infect mosquitoes and assess the ability of mutants to survive in mosquitoes and transmit the parasite.

Summer Intern – Manuel Llinás Lab 6/2012 – 9/2012
Princeton University

• Learned to clone plasmids for protein expression in E. coli
• Expressed and purified DNA binding domains of malaria transcription factors AP2 (Apetala 2 proteins) which regulate stages of the parasite life cycle
• Worked to define specific DNA binding sequences for each transcription factor

SELECTED PUBLICATIONS (4 of 20)

• Nasamu AS, Glushakova S, Russo I, Vaupel B, Oksman A, Kim AS, Fremont DH, Tolia N, Beck JR, Meyers MJ, Niles JC, Zimmerberg J, Goldberg DE. Plasmepsins IX and X are essential and druggable mediators of malaria parasite egress and invasion. Science. 2017 Oct 27;358(6362):518-522. doi: 10.1126/science.aan1478.
• Polino AJ*, Nasamu AS*, Niles JC, Goldberg DE. Assessment of Biological Role and Insight into Druggability of the Plasmodium falciparum Protease Plasmepsin V. ACS Infect Dis. 2020 Apr 10;6(4):738-746. doi: 10.1021/acsinfecdis.9b00460. Epub 2020 Feb 27. *These authors contributed equally
• Nasamu AS, Falla A, Pasaje CFA, Wall BA, Wagner JC, Ganesan SM, Goldfless SJ, Niles JC. An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum. Sci Rep. 2021 Jan 11;11(1):342. doi: 10.1038/s41598-020-77644-4.
• Mukherjee S*, Nasamu AS*, Rubiano KC, Goldberg DE. Activation of the Plasmodium Egress Effector Subtilisin-Like Protease 1 Is Mediated by Plasmepsin X Destruction of the Prodomain. mBio. 2023 Apr 25;14(2):e0067323. doi: 10.1128/mbio.00673-23. Epub 2023 Apr 10. *These authors contributed equally

Show All Publications

• Blackwell AM, Jami-Alahmadi Y, Nasamu AS, Kudo S, Senoo A, Slam C, Tsumoto K, Wohlschlegel JA, Manuel Martinez Caaveiro J, Goldberg DE, Sigala PA. Malaria parasites require a divergent heme oxygenase for apicoplast gene expression and biogenesis. Elife. 2024 Dec 11;13. doi: 10.7554/eLife.100256.
• Blackwell AM, Jami-Alahmadi Y, Nasamu AS, Kudo S, Senoo A, Slam C, Tsumoto K, Wohlschlegel JA, Caaveiro JMM, Goldberg DE, Sigala PA. Malaria parasites require a divergent heme oxygenase for apicoplast gene expression and biogenesis. bioRxiv. 2024 Oct 13;. doi: 10.1101/2024.05.30.596652.
• Mukherjee S, Nasamu AS, Rubiano KC, Goldberg DE. Activation of the Plasmodium Egress Effector Subtilisin-Like Protease 1 Is Mediated by Plasmepsin X Destruction of the Prodomain. mBio. 2023 Apr 25;14(2):e0067323. doi: 10.1128/mbio.00673-23. Epub 2023 Apr 10.
• Bopp S, Pasaje CFA, Summers RL, Magistrado-Coxen P, Schindler KA, Corpas-Lopez V, Yeo T, Mok S, Dey S, Smick S, Nasamu AS, Demas AR, Milne R, Wiedemar N, Corey V, Gomez-Lorenzo MG, Franco V, Early AM, Lukens AK, Milner D, Furtado J, Gamo FJ, Winzeler EA, Volkman SK, Duffey M, Laleu B, Fidock DA, Wyllie S, Niles JC, Wirth DF. Potent acyl-CoA synthetase 10 inhibitors kill Plasmodium falciparum by disrupting triglyceride formation. Nat Commun. 2023 Mar 16;14(1):1455. doi: 10.1038/s41467-023-36921-2.
• Mukherjee S, Nasamu AS, Rubiano K, Goldberg DE. Activation of the Plasmodium egress effector subtilisin-like protease 1 is achieved by plasmepsin X destruction of the propiece. bioRxiv. 2023 Jan 14;. doi: 10.1101/2023.01.13.524002.
• Cárdenas P, Esherick LY, Chambonnier G, Dey S, Turlo CV, Nasamu AS, Niles JC. GeneTargeter: Automated In Silico Design for Genome Editing in the Malaria Parasite, Plasmodium falciparum. CRISPR J. 2022 Feb;5(1):155-164. doi: 10.1089/crispr.2021.0069.
• Nasamu AS, Falla A, Pasaje CFA, Wall BA, Wagner JC, Ganesan SM, Goldfless SJ, Niles JC. An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum. Sci Rep. 2021 Jan 11;11(1):342. doi: 10.1038/s41598-020-77644-4.
• Nasamu AS, Polino AJ, Istvan ES, Goldberg DE. Malaria parasite plasmepsins: More than just plain old degradative pepsins. J Biol Chem. 2020 Jun 19;295(25):8425-8441. doi: 10.1074/jbc.REV120.009309. Epub 2020 May 4. Review.
• Polino AJ, Nasamu AS, Niles JC, Goldberg DE. Assessment of Biological Role and Insight into Druggability of the Plasmodium falciparum Protease Plasmepsin V. ACS Infect Dis. 2020 Apr 10;6(4):738-746. doi: 10.1021/acsinfecdis.9b00460. Epub 2020 Feb 27.
• Gupta A, Bokhari AAB, Pillai AD, Crater AK, Gezelle J, Saggu G, Nasamu AS, Ganesan SM, Niles JC, Desai SA. Complex nutrient channel phenotypes despite Mendelian inheritance in a Plasmodium falciparum genetic cross. PLoS Pathog. 2020 Feb;16(2):e1008363. doi: 10.1371/journal.ppat.1008363. eCollection 2020 Feb.
• Meyers MJ, Liu J, Xu J, Leng F, Guan J, Liu Z, McNitt SA, Qin L, Dai L, Ma H, Adah D, Zhao S, Li X, Polino AJ, Nasamu AS, Goldberg DE, Liu X, Lu Y, Tu Z, Chen X, Tortorella MD. 4-Aryl Pyrrolidines as a Novel Class of Orally Efficacious Antimalarial Agents. Part 1: Evaluation of 4-Aryl- N-benzylpyrrolidine-3-carboxamides. J Med Chem. 2019 Apr 11;62(7):3503-3512. doi: 10.1021/acs.jmedchem.8b01972. Epub 2019 Mar 22.
• Singh S, Rajendran V, He J, Singh AK, Achieng AO, Vandana, Pant A, Nasamu AS, Pandit M, Singh J, Quadiri A, Gupta N, Poonam, Ghosh PC, Singh BK, Narayanan L, Kempaiah P, Chandra R, Dunn BM, Pandey KC, Goldberg DE, Singh AP, Rathi B. Fast-Acting Small Molecules Targeting Malarial Aspartyl Proteases, Plasmepsins, Inhibit Malaria Infection at Multiple Life Stages. ACS Infect Dis. 2019 Feb 8;5(2):184-198. doi: 10.1021/acsinfecdis.8b00197. Epub 2019 Jan 2.
• Garten M, Nasamu AS, Niles JC, Zimmerberg J, Goldberg DE, Beck JR. EXP2 is a nutrient-permeable channel in the vacuolar membrane of Plasmodium and is essential for protein export via PTEX. Nat Microbiol. 2018 Oct;3(10):1090-1098. doi: 10.1038/s41564-018-0222-7. Epub 2018 Aug 27.
• Glushakova S, Beck JR, Garten M, Busse BL, Nasamu AS, Tenkova-Heuser T, Heuser J, Goldberg DE, Zimmerberg J. Rounding precedes rupture and breakdown of vacuolar membranes minutes before malaria parasite egress from erythrocytes. Cell Microbiol. 2018 Oct;20(10):e12868. doi: 10.1111/cmi.12868. Epub 2018 Jul 10.
• Nasamu AS, Glushakova S, Russo I, Vaupel B, Oksman A, Kim AS, Fremont DH, Tolia N, Beck JR, Meyers MJ, Niles JC, Zimmerberg J, Goldberg DE. Plasmepsins IX and X are essential and druggable mediators of malaria parasite egress and invasion. Science. 2017 Oct 27;358(6362):518-522. doi: 10.1126/science.aan1478.
• Sidik SM, Huet D, Ganesan SM, Huynh MH, Wang T, Nasamu AS, Thiru P, Saeij JPJ, Carruthers VB, Niles JC, Lourido S. A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2.
• Ganesan SM, Falla A, Goldfless SJ, Nasamu AS, Niles JC. Synthetic RNA-protein modules integrated with native translation mechanisms to control gene expression in malaria parasites. Nat Commun. 2016 Mar 1;7:10727. doi: 10.1038/ncomms10727.
• Chalkiadaki A, Igarashi M, Nasamu AS, Knezevic J, Guarente L. Muscle-specific SIRT1 gain-of-function increases slow-twitch fibers and ameliorates pathophysiology in a mouse model of duchenne muscular dystrophy. PLoS Genet. 2014 Jul;10(7):e1004490. doi: 10.1371/journal.pgen.1004490. eCollection 2014 Jul.

Show Selected Publications

TECHNICAL EXPERTISE

Molecular Biology, Genome Engineering and Microbiology

• Primer design, DNA assembly (Gibson, InFusion, Gateway, restriction/ligation, CRISPR/Cas9, CRISPR/RNP) in mammalian cells and parasites, cloning and genotyping
• Designing and constructing plasmids (using Benchling, Vector NTI, SnapGene)
• Mammalian cell transfection
• Using various genome engineering methods (spCRISPR/Cas9, eSpCas9, Cpf1, Bxb1, phlc3, etc.) for the development of stable transgenic cell lines (parasites and mammalian cells)
• Recombinant protein expression and purification
• DNA/RNA isolation, qRT-PCR, ddPCR, library preparation for Next Generation Sequencing
• Experience with a range of culture methods (E. coli, yeast, and Toxoplasma gondii)

Cell Biology and Biochemistry

• Mammalian cell culture, blood-borne disease cell culture (Plasmodium falciparum)
• Expertise in protein expression and characterization (SDS-PAGE, Western blot, immunoassays/ELISA, immunohistochemistry, electrophoretic mobility shift assays)
• Primary mammalian cell culture, passaging, and harvesting
• Flow cytometry (FACS) with fluorescent tags or antibodies
• Cell isolation (Ficoll separation), stage-specific parasite separation by MACS
• Immunoprecipitation, Chromatin-IP (ChIP), TUNEL assays.
• Light and fluorescence microscopy
• Generating and maintaining stable genetically engineered Toxoplasma gondii parasites in human cells
• Enzymology - isolation and purification of enzymes, enzyme kinetics, measuring activity, inhibitor assays

INVITED TALKS, ABSTRACTS AND POSTERS

• Invited speaker at the University of Georgia Center for Tropical & Emerging Global Diseases (2021). “The roles of Plasmepsins IX and X in malaria biology.”
• Invited speaker at the 66th Annual Meeting of the American Society for Tropical Medicine and Health (2017). “Plasmepsins IX and X are essential and druggable mediators of malaria parasite egress and invasion.”
• Invited speaker at the 28th Annual Molecular Parasitology Meeting (2017). “Plasmepsins IX and X are essential and druggable mediators of malaria parasite egress and invasion.”
• Invited speaker at the 5th Annual Global Health and Infectious Disease Trainee Oral Symposium and Conference (2017). “Plasmepsins IX and X are essential and druggable mediators of malaria parasite egress and invasion.”
• Invited speaker at the MIT NIH P50 Synthetic Biology Quarterly meetings (March 2015, June 2014, Oct. 2014, Dec. 2014). “Improved Donor Vector Assembly for Genome Editing Applications in Plasmodium falciparum.”
• Abstract and poster at the Molecular Parasitology Meeting (2015). “A robust genetic toolkit to explore novel genes in malaria parasites.”

AWARDS AND SCHOLARSHIPS

• American Heart Association Predoctoral Fellowship (2018 – 2019)
• Best Talk, Molecular Parasitology Meeting (2017)
• Media coverage: Podcast – This Week in Parasitism, Episode 143: http://www.microbe.tv/twip/twip-143/