Mendelian segregation and high recombination rates facilitate genetic analyses in Cryptosporidium parvum.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101239074 Publication Model: eCollection Cited Medium: Internet ISSN: 1553-7404 (Electronic) Linking ISSN: 15537390 NLM ISO Abbreviation: PLoS Genet Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science, c2005-

Cryptosporidium parvum*/genetics ; Oocysts* ; Recombination, Genetic* ; Cryptosporidiosis*/parasitology ; Cryptosporidiosis*/genetics ; Meiosis*/genetics; Animals ; Mice ; Humans ; Receptors, Interferon/genetics ; Interferon gamma Receptor ; Chromosome Segregation/genetics ; Sporozoites/genetics ; Mice, Knockout ; Phenotype

Very little is known about the process of meiosis in the apicomplexan parasite Cryptosporidium despite the essentiality of sex in its life cycle. Most cell lines only support asexual growth of Cryptosporidium parvum (C. parvum), but stem cell derived intestinal epithelial cells grown under air-liquid interface (ALI) conditions support the sexual cycle. To examine chromosomal dynamics during meiosis in C. parvum, we generated two transgenic lines of parasites that were fluorescently tagged with mCherry or GFP on chromosomes 1 or 5, respectively. Infection of ALI cultures or Ifngr1-/- mice with mCherry and GFP parasites resulted in cross-fertilization and the formation of "yellow" oocysts, which contain 4 haploid sporozoites that are the product of meiosis. Recombinant oocysts from the F1 generation were purified and used to infect HCT-8 cultures, and phenotypes of the progeny were observed by microscopy. All possible phenotypes predicted by independent segregation were represented equally (~25%) in the population, indicating that C. parvum chromosomes exhibit a Mendelian inheritance pattern. The most common pattern observed from the outgrowth of single oocysts included all possible parental and recombinant phenotypes derived from a single meiotic event, suggesting a high rate of crossover. To estimate the frequency of crossover, additional loci on chromosomes 1 and 5 were tagged and used to monitor intrachromosomal crosses in Ifngr1-/- mice. Both chromosomes showed a high frequency of crossover compared to other apicomplexans with map distances (i.e., 1% recombination) of 3-12 kb. Overall, a high recombination rate may explain many unique characteristics observed in Cryptosporidium spp. such as high rates of speciation, wide variation in host range, and rapid evolution of host-specific virulence factors.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 Kimball et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Update of: bioRxiv. 2024 Feb 02:2024.02.02.578536. doi: 10.1101/2024.02.02.578536. (PMID: 38352509)

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R01 AI175150 United States AI NIAID NIH HHS

0 (Receptors, Interferon)
0 (Interferon gamma Receptor)

Date Created: 20240617 Date Completed: 20240628 Latest Revision: 20240705

20240705

PMC11213348

10.1371/journal.pgen.1011162

38885280