Cyst induction by intracellular Mg2+/Ca2+ injection and its implications for optimizing Mg2+ conditions for future CRISPR delivery in <italic style="font-style: italic">Alexandrium</italic> <italic style="font-style: italic">catenella</italic>

Zhenfan CHEN; Chang GUO; Huixia GENG; Fanzhou KONG; Qingchun ZHANG; Rencheng YU

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Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella

Updated：2026-04-07

- Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella
- Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella
- 海洋湖沼学报(英文) 2026年页码：1-10
- Affiliations：
  
  1.Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China
  2.Laboratory for Marine Ecology and Environmental Science, Qingdao Marine Science and Technology Center, Qingdao 266071, China
  3.University of Chinese Academy of Sciences, Beijing 100049, China
- Author bio：
  
  rcyu@qdio.ac.cn
- Funds：
  
  the National Key R&D Programs from the Ministry of Science and Technology (MoST) of China(2022YFC3105300;2023YFE0106200);the Taishan Scholarship of Professor Rencheng YU, the China Postdoctoral Science Foundation(2023M733523);the Shandong Provincial Natural Science Foundation(ZR2023QD132);the Qingdao Postdoctoral Applied Research Project(QDBSH20230202008)
- DOI：
  中图分类号：
- 收稿：2025-05-19，
  
  网络首发：2026-04-07，
- Accepted：
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Zhenfan CHEN, Chang GUO, Huixia GENG, 等. Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella[J/OL]. 海洋湖沼学报(英文), 2026,1-10.

CHEN Zhenfan,GUO Chang,GENG Huixia,et al.Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella[J].Journal of Oceanology and Limnology,
Zhenfan CHEN, Chang GUO, Huixia GENG, 等. Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella[J/OL]. 海洋湖沼学报(英文), 2026,1-10. DOI：

CHEN Zhenfan,GUO Chang,GENG Huixia,et al.Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella[J].Journal of Oceanology and Limnology, DOI：.

摘要

Abstract

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas system is a powerful tool for gene editing

yet it has never been successfully applied to edit genes in dinoflagellates. In CRISPR-Cas-mediated gene editing

magnesium ions (Mg

) are necessary to maintain catalytic activity. Some studies

however

found that Mg

and calcium (Ca

) ions could modulate flagellar motility and cyst formation of unicellular flagellates

thus affecting their physiology. Here

a single-cell microinjection method was adapted to deliver Mg

/Ca

into cells of the dinoflagellate

Alexandrium

catenella

to test the dose-dependent effects on cyst formation and germination. It was found that Mg

/Ca

treatments

https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=103448613&type=

https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=103448600&type=

1.60866666

2.28600001

50 mmol/L significantly enhanced cyst survival and germination rates on day 1 compared to those of higher concentrations. In contrast

treatments with high concentrations (

https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=103448601&type=

https://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=103448588&type=

1.60866666

2.28600001

250 mmol/L) of Mg

/Ca

triggered irreversible dormancy transitions

accompanied by increased mortality and prolonged germination time. The treatment with 10-mmol/L Mg

achieved an optimal balance between CRISPR-Cas nuclease activity and cyst germination. This work provides the first experimental assessment of ion-induced cyst physiology using single-cell microinjection in dinoflagellates

and identifies Mg

concentrations that are physiologically compatible with microinjection. These findings establish the fundamental conditions for future microinjection-based CRISPR delivery in

catenella

关键词

Keywords

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Cyst induction by intracellular Mg2+/Ca2+ injection and its implications for optimizing Mg2+ conditions for future CRISPR delivery in Alexandrium catenella

Cyst induction by intracellular Mg2+/Ca2+ injection and its implications for optimizing Mg2+ conditions for future CRISPR delivery in Alexandrium catenella

DOI：

摘要

Abstract

关键词

Keywords

references

Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella

Cyst induction by intracellular Mg²⁺/Ca²⁺ injection and its implications for optimizing Mg²⁺ conditions for future CRISPR delivery in Alexandrium catenella