Show simple item record

dc.contributor.author
Knuepfer, Ellen
dc.contributor.author
Napiorkowska, Marta
dc.contributor.author
van Ooij, Christiaan
dc.contributor.author
Holder, Anthony A.
dc.date.accessioned
2017-11-21T13:20:16Z
dc.date.available
2017-10-06T02:14:17Z
dc.date.available
2017-11-21T13:20:16Z
dc.date.issued
2017-06-20
dc.identifier.issn
2045-2322
dc.identifier.other
10.1038/s41598-017-03984-3
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/190564
dc.identifier.doi
10.3929/ethz-b-000190564
dc.description.abstract
Successful establishment of CRISPR/Cas9 genome editing technology in Plasmodium spp. has provided a powerful tool to transform Plasmodium falciparum into a genetically more tractable organism. Conditional gene regulation approaches are required to study the function of gene products critical for growth and erythrocyte invasion of blood stage parasites. Here we employ CRISPR/Cas9 to facilitate use of the dimerisable Cre-recombinase (DiCre) that is frequently used to mediate the excision and loss of loxP-flanked DNA sequences in a rapamycin controlled manner. We describe novel CRISPR/Cas9 transfection plasmids and approaches for the speedy, stable and marker-free introduction of transgenes encoding the DiCre recombinase into genomic loci dispensable for blood stage development. Together these plasmids form a toolkit that will allow the rapid generation of transgenic DiCre-expressing P. falciparum lines in any genetic background. Furthermore, the newly developed 3D7-derived parasite lines, constitutively and stably expressing DiCre, generated using this toolkit will prove useful for the analysis of gene products. Lastly, we introduce an improved treatment protocol that uses a lower rapamycin concentration and shorter treatment times, leading to loxP-guided recombination with close to 100% efficiency within the same replication cycle.
en_US
dc.format
application/pdf
dc.language.iso
en
en_US
dc.publisher
Nature Publishing Group
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.title
Generating conditional gene knockouts in Plasmodium - a toolkit to produce stable DiCre recombinase-expressing parasite lines using CRISPR/Cas9
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
ethz.journal.title
Scientific Reports
ethz.journal.volume
7
en_US
ethz.journal.abbreviated
Sci Rep
ethz.pages.start
3881
en_US
ethz.size
12 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.status
published
en_US
ethz.date.deposited
2017-10-06T02:14:30Z
ethz.source
WOS
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2017-11-21T13:20:20Z
ethz.rosetta.lastUpdated
2021-02-14T20:35:38Z
ethz.rosetta.versionExported
true
ethz.COinS
ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.atitle=Generating%20conditional%20gene%20knockouts%20in%20Plasmodium%20-%20a%20toolkit%20to%20produce%20stable%20DiCre%20recombinase-expressing%20parasite%20lines%20using%20CRI&rft.jtitle=Scientific%20Reports&rft.date=2017-06-20&rft.volume=7&rft.spage=3881&rft.issn=2045-2322&rft.au=Knuepfer,%20Ellen&Napiorkowska,%20Marta&van%20Ooij,%20Christiaan&Holder,%20Anthony%20A.&rft.genre=article&rft_id=info:doi/10.1038/s41598-017-03984-3&
 Search print copy at ETH Library

Files in this item

Thumbnail

Publication type

Show simple item record