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. 2024 Mar 21;16(4):723–754. doi: 10.1038/s44321-024-00057-7

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) PfLARC2 was created by first deleting Pf Mei2 on chromosome 6 and then Pf LINUP on chromosome 12 using plasmids pFC-yFCU-Plasmei2 and pFC-yFCU-LINUP, respectively. Sequential positive and negative selection with WR99210 and 5FC resulted in marker-free PfLARC2 free of extraneous DNA. Primers used for confirming parasite genotypes are shown as letters. Expected amplicon sizes for WT and PfLARC2 and the pFC plasmid are indicated in the schematic. Agarose gel electrophoresis to confirm, (B) gene deletion of PlasMei2, (C) gene deletion of LINUP, and (D) lack of plasmid in three PfLARC2 clones F7, C12, and B7, respectively. Source data are available online for this figure.