Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9

Methods Mol Biol. 2020:2052:219-228. doi: 10.1007/978-1-4939-9748-0_13.

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: CRISPR; Cas9; Genetics; Transfection; Transgenics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Associated Protein 9*
  • CRISPR-Cas Systems*
  • Cell Line
  • Cryptosporidiosis / parasitology
  • Cryptosporidium parvum / genetics*
  • Cryptosporidium parvum / growth & development
  • Cryptosporidium parvum / isolation & purification
  • Genetic Techniques
  • Genetic Vectors
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism
  • Mice
  • Oocysts / genetics
  • Oocysts / growth & development
  • Oocysts / isolation & purification
  • Transfection / instrumentation
  • Transfection / methods*
  • Transgenes
  • Workflow

Substances

  • Luciferases
  • CRISPR-Associated Protein 9