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鱿鱼原代细胞的分离、培养及转染方案。

Protocol for the isolation, culture, and transfection of squid primary cells.

作者信息

Kim Yoonji, Tanner Hailey M, Rosenthal Joshua J C, Brangwynne Clifford P

机构信息

Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA; Omenn-Darling Bioengineering Institute, Princeton University, Princeton, NJ 08544, USA.

Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA.

出版信息

STAR Protoc. 2025 Aug 5;6(3):103994. doi: 10.1016/j.xpro.2025.103994.

DOI:10.1016/j.xpro.2025.103994
PMID:40773350
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12347879/
Abstract

We present a protocol for the isolation, culture, and transfection of squid primary cells from various tissues and ages by including steps for squid dissection, cell isolation, maintenance in culture, and expression of exogenous genes. We also highlight life-stage tractability in Euprymna berryi squids for future aging studies. This protocol enables molecular- and cellular-level investigations into processes specific to squids, including their complex behaviors, rapid color change mechanisms, and RNA editing capabilities, with broader implications in basic cell physiology.

摘要

我们提出了一种从不同组织和年龄的鱿鱼中分离、培养和转染原代细胞的方案,其中包括鱿鱼解剖、细胞分离、培养维持以及外源基因表达等步骤。我们还强调了柏氏艾氏鱿鱼在生命阶段方面的易处理性,以便未来进行衰老研究。该方案能够在分子和细胞水平上对鱿鱼特有的过程进行研究,包括它们复杂的行为、快速的颜色变化机制和RNA编辑能力,对基础细胞生理学具有更广泛的意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/e26b0dc18825/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/cc3d8b36c7a4/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/b521d3bd1242/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/fc851197410f/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/1f58fcee0f8a/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/e26b0dc18825/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/cc3d8b36c7a4/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/b521d3bd1242/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/fc851197410f/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/1f58fcee0f8a/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/135a/12347879/e26b0dc18825/gr7.jpg

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本文引用的文献

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Nat Ecol Evol. 2025 Jun 6. doi: 10.1038/s41559-025-02720-9.
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High-level RNA editing diversifies the coleoid cephalopod brain proteome.高水平的 RNA 编辑使头足类动物大脑蛋白质组多样化。
Brief Funct Genomics. 2023 Nov 17;22(6):525-532. doi: 10.1093/bfgp/elad034.
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Creation of an albino squid line by CRISPR-Cas9 and its application for in vivo functional imaging of neural activity.
通过 CRISPR-Cas9 技术创建白化鱿鱼品系及其在体内神经活动功能成像中的应用。
Curr Biol. 2023 Jul 10;33(13):2774-2783.e5. doi: 10.1016/j.cub.2023.05.066. Epub 2023 Jun 20.
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Gene delivery available in molluscan cells by strong promoter discovered from bivalve-infectious virus.从双壳类传染性病毒中发现的强启动子可用于软体动物细胞的基因传递。
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