{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T04:40:33Z","timestamp":1781844033563,"version":"3.54.5"},"reference-count":39,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,1,8]],"date-time":"2021-01-08T00:00:00Z","timestamp":1610064000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81630063"],"award-info":[{"award-number":["81630063"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>Studies of molecular mechanisms and related gene functions have long been restricted by limited genome editing technologies in malaria parasites. Recently, a simple and effective genome editing technology, the CRISPR\/Cas (clustered regularly interspaced short palindromic repeats\/CRISPR-associated) system, has greatly facilitated these studies in many organisms, including malaria parasites. However, due to the special genome feature of malaria parasites, the manipulation and gene editing efficacy of the CRISPR\/Cas system in this pathogen need to be improved, particularly in the human malaria parasite, <jats:italic>Plasmodium falciparum<\/jats:italic>. Herein, based on the CRISPR\/Cas9 system, we developed an integrating strategy to generate a Cas9i system, which significantly shortened the time for generation of transgenic strains in <jats:italic>P. falciparum<\/jats:italic>. Moreover, with this Cas9i system, we have successfully achieved multiplexed genome editing (mutating or tagging) by a single-round transfection in <jats:italic>P. falciparum<\/jats:italic>. In addition, we for the first time adapted AsCpf1 (<jats:italic>Acidaminococcus<\/jats:italic> sp. Cpf1), an alternative to Cas9, into <jats:italic>P. falciparum<\/jats:italic> parasites and examined it for gene editing. These optimizations of the CRISPR\/Cas system will further facilitate the mechanistic research of malaria parasites and contribute to eliminating malaria in the future.<\/jats:p>","DOI":"10.3389\/fmicb.2020.625862","type":"journal-article","created":{"date-parts":[[2021,1,8]],"date-time":"2021-01-08T04:35:17Z","timestamp":1610080517000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":15,"title":["Optimization of CRISPR\/Cas System for Improving Genome Editing Efficiency in Plasmodium falciparum"],"prefix":"10.3389","volume":"11","author":[{"given":"Yuemeng","family":"Zhao","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Changhong","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaobai","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cizhong","family":"Jiang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Ding","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Shen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingfeng","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,1,8]]},"reference":[{"key":"B2","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1038\/nature12876","article-title":"A molecular marker of artemisinin-resistant Plasmodium falciparum malaria.","volume":"505","author":"Ariey","year":"2014","journal-title":"Nature"},{"key":"B3","doi-asserted-by":"publisher","first-page":"933","DOI":"10.1038\/nbt.3659","article-title":"Applications of CRISPR technologies in research and beyond.","volume":"34","author":"Barrangou","year":"2016","journal-title":"Nat. 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