TY - JOUR
T1 - Genetic crosses within and between species of Cryptosporidium
AU - Shaw, Sebastian
AU - Cohn, Ian S.
AU - Baptista, Rodrigo P.
AU - Xia, Guoqin
AU - Melillo, Bruno
AU - Agyabeng-Dadzie, Fiifi
AU - Kissinger, Jessica C.
AU - Striepen, Boris
N1 - Funding Information:
This work was supported in part by the NIH with grants to B.S. (R01AI112427 and R01AI127798), J.C.K. and Travis Glenn (R01AI148667), a fellowship to I.S.C. (F30AI169744), support to F.A.-D. (T32GM142623), and fellowships from the Swiss NSF to S.S. (P2BEP3_191774 and P500PB_211097). We thank Chyi Hsieh for sharing C. tyzzeri oocysts, and Abhijit Kundu (TCGLS) for small molecule synthesis, and Travis Glenn for help with single oocyst sequencing and mentorship of F.A.-D.
Funding Information:
ACKNOWLEDGMENTS. This work was supported in part by the NIH with grants to B.S. (R01AI112427 and R01AI127798), J.C.K. and Travis Glenn (R01AI148667), a fellowship to I.S.C. (F30AI169744), support to F.A.-D. (T32GM142623), and fellowships from the Swiss NSF to S.S. (P2BEP3_191774 and P500PB_211097). We thank Chyi Hsieh for sharing C. tyzzeri oocysts, and Abhijit Kundu (TCGLS) for small molecule synthesis, and Travis Glenn for help with single oocyst sequencing and mentorship of F.A.-D.
Publisher Copyright:
Copyright © 2023 the Author(s). Published by PNAS.
PY - 2023/12/26
Y1 - 2023/12/26
N2 - Parasites and their hosts are engaged in rapid coevolution that balances competing mechanisms of virulence, resistance, and evasion. This often leads to host specificity, but genomic reassortment between different strains can enable parasites to jump host barriers and conquer new niches. In the apicomplexan parasite
Cryptosporidium genetic exchange has been hypothesized to play a prominent role in adaptation to humans. The sexual lifecycle of the parasite provides a potential mechanism for such exchange; however, the boundaries of
Cryptosporidium sex are currently undefined. To explore this experimentally, we established a model for genetic crosses. Drug resistance was engineered using a mutated phenylalanyl tRNA synthetase gene and marking strains with this and the previously used Neo transgene enabled selection of recombinant progeny. This is highly efficient, and genomic recombination is evident and can be continuously monitored in real time by drug resistance, flow cytometry, and PCR mapping. Using this approach multiple loci can now be modified with ease. We demonstrate that essential genes can be ablated by crossing a Cre recombinase driver strain with floxed strains. We further find that genetic crosses are also feasible between species. Crossing
C. parvum, a parasite of cattle and humans, and
C. tyzzeri a mouse parasite resulted in progeny with a recombinant genome derived from both species that continues to vigorously replicate sexually. These experiments have important fundamental and translational implications for the evolution of
Cryptosporidium and open the door to reverse- and forward- genetic analysis of parasite biology and host specificity.
AB - Parasites and their hosts are engaged in rapid coevolution that balances competing mechanisms of virulence, resistance, and evasion. This often leads to host specificity, but genomic reassortment between different strains can enable parasites to jump host barriers and conquer new niches. In the apicomplexan parasite
Cryptosporidium genetic exchange has been hypothesized to play a prominent role in adaptation to humans. The sexual lifecycle of the parasite provides a potential mechanism for such exchange; however, the boundaries of
Cryptosporidium sex are currently undefined. To explore this experimentally, we established a model for genetic crosses. Drug resistance was engineered using a mutated phenylalanyl tRNA synthetase gene and marking strains with this and the previously used Neo transgene enabled selection of recombinant progeny. This is highly efficient, and genomic recombination is evident and can be continuously monitored in real time by drug resistance, flow cytometry, and PCR mapping. Using this approach multiple loci can now be modified with ease. We demonstrate that essential genes can be ablated by crossing a Cre recombinase driver strain with floxed strains. We further find that genetic crosses are also feasible between species. Crossing
C. parvum, a parasite of cattle and humans, and
C. tyzzeri a mouse parasite resulted in progeny with a recombinant genome derived from both species that continues to vigorously replicate sexually. These experiments have important fundamental and translational implications for the evolution of
Cryptosporidium and open the door to reverse- and forward- genetic analysis of parasite biology and host specificity.
KW - Apicomplexa
KW - diarrheal disease
KW - genetic cross
KW - parasite
KW - sex
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U2 - 10.1073/pnas.2313210120
DO - 10.1073/pnas.2313210120
M3 - Article
C2 - 37577700
SN - 0027-8424
VL - 121
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 1
M1 - e2313210120
ER -