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Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9

  • Adam Sateriale
  • Mattie Pawlowic
  • Sumiti Vinayak
  • Carrie Brooks
  • Boris StriepenEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2052)

Abstract

Cryptosporidium parvum can be reliably genetically manipulated using CRISPR/Cas9-driven homologous repair coupled to in vivo propagation within immunodeficient mice. Recent modifications have simplified the initial protocol significantly. This chapter will guide through procedures for excystation, transfection, infection, collection, and purification of transgenic Cryptosporidium parvum.

Keywords

Transfection Transgenics CRISPR Cas9 Genetics 

Notes

Acknowledgments

Our work on Cryptosporidium transgenesis was supported by grants from the National Institutes of Health, the Wellcome Trust, and the Bill & Melinda Gates Foundation to B.S. MCP was supported by NIH T32AI060546 training grant, and AS received F32AI116053 and K99AI37442 career fellowships from the National Institutes of Health.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Adam Sateriale
    • 1
  • Mattie Pawlowic
    • 2
  • Sumiti Vinayak
    • 3
  • Carrie Brooks
    • 4
  • Boris Striepen
    • 1
    Email author
  1. 1.Department of Pathobiology, School of Veterinary MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.Wellcome Trust Centre for Anti-Infectives ResearchUniversity of DundeeDundeeUK
  3. 3.Department of Pathobiology, College of Veterinary MedicineUniversity of Illinois Urbana-ChampaignUrbanaUSA
  4. 4.Center for Tropical and Emerging and Global DiseasesUniversity of GeorgiaAthensUSA

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