Show simple item record

dc.contributor.authorKanjee, Usheeren
dc.contributor.authorGrüring, Christofen
dc.contributor.authorChaand, Muditen
dc.contributor.authorLin, Kai-Minen
dc.contributor.authorEgan, Elizabethen
dc.contributor.authorManzo, Jaleen
dc.contributor.authorJones, Patrick Len
dc.contributor.authorYu, Tiffanyen
dc.contributor.authorBarker, Roberten
dc.contributor.authorWeekes, Michaelen
dc.contributor.authorDuraisingh, Manoj Ten
dc.date.accessioned2018-05-23T12:56:34Z
dc.date.available2018-05-23T12:56:34Z
dc.date.issued2017-10-19en
dc.identifier.issn0027-8424
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/276127
dc.description.abstractDuring malaria blood stage infections, Plasmodium parasites interact with the red blood cell (RBC) surface to enable invasion followed by intracellular proliferation. Critical factors involved in invasion have been identified using biochemical and genetic approaches including specific knockdowns of genes of interest from primary CD34+ hematopoietic stem cells (cRBCs). Here, we report the development of a robust in vitro culture system to produce RBCs that allow for generation of gene knockouts via CRISPR/Cas9 using the immortal JK-1 erythroleukemia line. JK-1 cells spontaneously differentiate, generating cells at different stages of erythropoiesis, including terminally differentiated nucleated RBCs that we term “jkRBCs”. A screen of small molecule epigenetic regulators identified several bromodomain-specific inhibitors that promote differentiation, and enable production of synchronous populations of jkRBCs. Global surface proteomic profiling revealed that jkRBCs express all known P. falciparum host receptors in a similar fashion to cRBCs and multiple P. falciparum strains invade jkRBCs at comparable levels to cRBCs and RBCs. Using CRISPR/Cas9 we deleted two host factors (BSG and CD44) for which no natural nulls exist. BSG interacts with the parasite ligand Rh5, a prominent vaccine candidate. A BSG knockout was completely refractory to parasite invasion in a strain-transcendent manner, confirming the essential role for BSG during invasion. CD44 was recently identified in an RNAi screen of blood group genes as a host factor for invasion, and we show that a CD44 knockout results in strain-transcendent reduction in invasion. Furthermore we demonstrate a functional interaction between these two determinants in mediating P. falciparum erythrocyte invasion.
dc.format.mediumPrint-Electronicen
dc.languageengen
dc.publisherNational Academy of Sciences
dc.subjectErythrocytesen
dc.subjectHematopoietic Stem Cellsen
dc.subjectCell Line, Tumoren
dc.subjectK562 Cellsen
dc.subjectHumansen
dc.subjectPlasmodium falciparumen
dc.subjectMalariaen
dc.subjectMalaria, Falciparumen
dc.subjectLeukemia, Erythroblastic, Acuteen
dc.subjectCarrier Proteinsen
dc.subjectProtozoan Proteinsen
dc.subjectAntigens, Protozoanen
dc.subjectLigandsen
dc.subjectProteomicsen
dc.subjectCell Differentiationen
dc.subjectEpigenesis, Geneticen
dc.subjectHost-Parasite Interactionsen
dc.subjectGene Knockout Techniquesen
dc.subjectCRISPR-Cas Systemsen
dc.subjectClustered Regularly Interspaced Short Palindromic Repeatsen
dc.subjectHyaluronan Receptorsen
dc.subjectBasiginen
dc.titleCRISPR/Cas9 knockouts reveal genetic interaction between strain-transcendent erythrocyte determinants of Plasmodium falciparum invasion.en
dc.typeArticle
prism.endingPageE9365
prism.issueIdentifier44en
prism.publicationDate2017en
prism.publicationNameProceedings of the National Academy of Sciences of the United States of Americaen
prism.startingPageE9356
prism.volume114en
dc.identifier.doi10.17863/CAM.23409
dcterms.dateAccepted2017-09-28en
rioxxterms.versionofrecord10.1073/pnas.1711310114en
rioxxterms.versionAM*
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2017-10-19en
dc.contributor.orcidKanjee, Usheer [0000-0002-3169-4557]
dc.contributor.orcidGrüring, Christof [0000-0001-8741-0747]
dc.contributor.orcidLin, Kai-Min [0000-0003-2109-1530]
dc.contributor.orcidWeekes, Michael [0000-0003-3196-5545]
dc.identifier.eissn1091-6490
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idWELLCOME TRUST (108070/Z/15/Z)


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record