Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9

Adam Sateriale, Mattie Pawlowic, Sumiti Vinayak, Carrie Brooks, Boris Striepen (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish
Title of host publicationCryptosporidium
EditorsJan R. Mead, Michael J. Arrowood
Place of PublicationNew York
PublisherHumana Press
Chapter13
Pages219-228
Number of pages10
ISBN (Electronic)9781493997480
ISBN (Print)9781493997473
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
PublisherSpringer Verlag
Volume2052
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Cryptosporidium parvum
Transfection
Infection

Keywords

  • CRISPR
  • Cas9
  • Genetics
  • Transfection
  • Transgenics

Cite this

Sateriale, A., Pawlowic, M., Vinayak, S., Brooks, C., & Striepen, B. (2020). Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9. In J. R. Mead, & M. J. Arrowood (Eds.), Cryptosporidium (pp. 219-228). (Methods in Molecular Biology; Vol. 2052). New York: Humana Press. https://doi.org/10.1007/978-1-4939-9748-0_13
Sateriale, Adam ; Pawlowic, Mattie ; Vinayak, Sumiti ; Brooks, Carrie ; Striepen, Boris. / Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9. Cryptosporidium. editor / Jan R. Mead ; Michael J. Arrowood. New York : Humana Press, 2020. pp. 219-228 (Methods in Molecular Biology).
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Sateriale, A, Pawlowic, M, Vinayak, S, Brooks, C & Striepen, B 2020, Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9. in JR Mead & MJ Arrowood (eds), Cryptosporidium. Methods in Molecular Biology, vol. 2052, Humana Press, New York, pp. 219-228. https://doi.org/10.1007/978-1-4939-9748-0_13

Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9. / Sateriale, Adam; Pawlowic, Mattie; Vinayak, Sumiti; Brooks, Carrie; Striepen, Boris (Lead / Corresponding author).

Cryptosporidium. ed. / Jan R. Mead; Michael J. Arrowood. New York : Humana Press, 2020. p. 219-228 (Methods in Molecular Biology; Vol. 2052).

Research output: Chapter in Book/Report/Conference proceedingChapter

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N1 - 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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AB - 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.

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KW - Cas9

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Sateriale A, Pawlowic M, Vinayak S, Brooks C, Striepen B. Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9. In Mead JR, Arrowood MJ, editors, Cryptosporidium. New York: Humana Press. 2020. p. 219-228. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9748-0_13