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HK-CRISPR-NUP205-mEGFP
Tissue
宫颈内口
Disease
腺癌
Metastatic site
原发肿瘤部位（宫颈管）
Synonyms
HK-CRISPR-NUP205-mEGFP #81

Documents

HK-CRISPR-NUP205-mEGFP

Cell line · Product number 301574

The HK-CRISPR-NUP205-mEGFP cell line is a genetically engineered human cell line designed to study nucleoporin 205 (NUP205) and its role in the nuclear po...

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01General information

Description

HK-CRISPR-NUP205-mEGFP 细胞系是一种基因工程人类细胞系，旨在研究核蛋白 205 (NUP205) 及其在核孔复合体中的作用。通过 CRISPR-Cas9 改造，用单体增强型绿色荧光蛋白（mEGFP）标记 NUP205，它能在活细胞中可视化和跟踪 NUP205，有助于研究核转运机制和核孔复合体动态。

NUP205 是核孔复合体的重要组成部分，负责调节细胞核与细胞质之间的分子运输。用 mEGFP 标记 NUP205 可让研究人员在荧光显微镜下实时观察其定位和行为，因此这种细胞系特别适用于研究核孔复合体的结构和功能及其在基因表达、RNA 处理和细胞周期中的作用。

HK-CRISPR-NUP205-mEGFP细胞系是研究核胞质转运机制和核孔复合体在细胞平衡中作用的有力工具。它对于探索核孔功能紊乱如何导致癌症和神经退行性疾病等疾病也很有价值，为促进我们对核转运及其对人类健康影响的理解提供了一个强大的模型。

Tissue
宫颈内口
Disease
腺癌
Metastatic site
原发肿瘤部位（宫颈管）
Applications
核孔复合体（NPC）支架生物学；NUP205支架核孔蛋白成像；mEGFP活细胞可视化；超分辨率显微镜；CRISPR敲入验证；核-胞质转运；NPC结构组织
Synonyms
  • HK-CRISPR-NUP205-mEGFP #81

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HeLa Kyoto line contains a CRISPR-engineered mEGFP fusion at the NUP205 locus for scaffold-level nuclear pore research. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
具有镶嵌石形状的上皮样细胞
Cell type
上皮细胞

04Identifiers

Citation
HK-CRISPR-NUP205-mEGFP（Cytion 目录号 301574）
NCBI TaxID
9606
Cellosaurus
CVCL_UR49
Depositor
艾伦伯格实验室（EMBL）

05Expression and susceptibility

Products
EGFP（增强型绿色荧光蛋白）

06Handling and culture

Culture medium
DMEM，w：4.5 克/升葡萄糖，w：4 毫摩尔 L-谷氨酰胺，w：3.7 克/升 NaHCO3，w：1.0 毫摩尔丙酮酸钠（Cytion 编号 820300a）。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基，用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS，T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞，T25 烧瓶用 1-2 毫升，T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟，使其分离。孵育后，用 10 毫升培养基轻轻混合细胞使其重悬，然后用 300xg 离心 3 分钟。弃去上清液，用新鲜培养基重悬细胞，并将其转移到已装有新鲜培养基的新烧瓶中。
Fluid renewal
每周 2 至 3 次
Thawing and culturing

  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染，每天都要对细胞培养物进行目视检查。

Freeze medium
我们使用完全生长培养基（包括 FBS）+10% DMSO 或 CM-1（Cytion 商品目录编号 800100）作为冷冻保存培养基，以获得足够的解冻后存活率，CM-1（Cytion 商品目录编号 800100）含有优化的渗透保护剂和代谢稳定剂，可提高恢复能力并减少冷冻引起的应激。
Storage conditions

如需长期保存，可将样品瓶放入气相液氮中，温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

Shipping conditions

冷冻保存的细胞系用干冰装运，并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后，应立即检查容器，并立即将小瓶转移到适当的储藏室。

08Donor

Donor age
30年
Ethnicity
非洲裔美国人