Community Resources

Over the last 7 years, the MCC has generated and phenotyped 4 P. falciparum genetic crosses, all with multiple biological replicates, resulting in hundreds of unique recombinant progeny. These crosses have been used for identifying genetic determinants of resistance to frontline antimalarial drugs, nutritional requirements and metabolism, parasite fitness, single-cell transcriptomics, and more.

The generation of these crosses relies on our novel human hepatocyte-liver chimeric mouse model (the FRG huHep mouse). Anopheles stephensi mosquitoes are fed on P. falciparium cultures adjusted to contain equal portions of gametocytes from each parental parasite. Recombinant sporozoites are injected into the FRG huHep mice 6 days after inoculation in the mosquito. During the transition from liver to blood stages, the mice are injected with human red blood cells before being exsanguinated to recover bulk pool of recombinant parasites. Individual parasites are isolated through limited dilution cloning and genotyped via Illumina sequencing.

A flowchart diagrams the experimental genetic cross of *Plasmodium falciparum* malaria parasites. Starting with in vitro blood stages in week one, the process moves to in vivo mosquito in weeks 2-3, then an FRG huHep/huRBC mouse in weeks 4-5. Weeks 6-12 show the return to in vitro blood stages, cloning and expansion. Finally, weeks 12-16 involve F1 progeny genotyping, culminating in a genetic map in weeks 17-18. Images and diagrams illustrate each stage of this cross.

Over the last 7 years, the MCC has generated and phenotyped 4 P. faciparum genetic crosses, all with multiple biological replicates, resulting in hundreds of unique recombinant progeny. Sequence data for these crosses can be found on the NIH sequence read archive (SRA) website (see pages of individual crosses for accession numbers).

Parental Strains Number of Unique Recombinants MMC Experiments
NF54 (Africa)
Drug Sensitive
NHP4026 (Asia)
CQ Resistant
270 Lifecycle, drug, and nutrient bulk segregant analysis; organelle inheritance, outcrossing, drug and fitness phenotyping
MKK2835 (Asia)
ART-S; CQ-R
NHP1337 (Asia)
ART-R; CQ-R
60 Lifecycle and drug bulk segregant analysis; drug phenotyping
Mal31 (Africa)
Drug Sensitive
KH004 (Asia)
ART-R; CQ-R; PPQ-R
104 Drug bulk segregant analysis; drug and fitness phenotyping
NF54 (Africa)
Drug Sensitive
NHP1337 (Asia)
ART-R; CQ-R
Bulk pool only Drug bulk segregant analysis
NF54 (Africa)
Drug Sensitive
7G8 (South America)
CQ-R
  Recombination rate. Mouse model proof of concept
NF54 (Africa)
Drug Sensitive
GB4 (Africa)
CQ-R
  Recombination rate. Mouse model proof of concept

To inquire about progeny from any of our genetic crosses, please email mcc@nd.edu