Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9
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.
KeywordsTransfection Transgenics CRISPR Cas9 Genetics
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.
- 3.Abrahamsen MS, Templeton TJ, Enomoto S, Abrahante JE, Zhu G, Lancto CA, Deng M, Liu C, Widmer G, Tzipori S, Buck GA, Xu P, Bankier AT, Dear PH, Konfortov BA, Spriggs HF, Iyer L, Anantharaman V, Aravind L, Kapur V (2004) Complete genome sequence of the apicomplexan, Cryptosporidium parvum. Science 304(5669):441–445. https://doi.org/10.1126/science.1094786CrossRefPubMedGoogle Scholar