Geneticin, G-418 Sulfate
(Synonyms: G-418硫酸盐,Antibiotic G418) 目录号 : GC17427
An aminoglycoside antibiotic
Cas No.:108321-42-2
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
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- Purity: >98.00%
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- Datasheet
| Cell experiment [1]: | |
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Cell lines |
Sheep skin fibroblasts, SSFs |
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Preparation Method |
The SSFs cultured in vitro were treated with different concentrations of single antibiotic Geneticin, G-418 Sulfate. After 12 days of treatment, the SSFs were digested, and the number of dead and alive cells was counted after placental blue staining to determine the lowest lethal dose of Geneticin, G-418 Sulfate leading to the death of all SSFs. |
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Reaction Conditions |
100-200ug/ml Geneticin, G-418 Sulfate for 1 day |
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Applications |
Treatment with 100 µg/mL Geneticin, G-418 Sulfate for 12 days did not lead to the death of all SSFs, but still (44.7 0.05)% of SSFs survived.Continuous treatment with 200 µg/mL Geneticin, G-418 Sulfate for 12 days can lead to the death of all SSFs. Therefore, the minimum lethal concentration of Geneticin, G-418 Sulfate leading to the death of SSFs is 200 µg/mL. |
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References: [1]: Wu Xian, JING Qian-ge, et,al.Study on the resistance of sheep skin fibroblasts to G418 and Blasticidin S [J].Genomics and applied biology,2019,38(03):1006-1011. |
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Geneticin, G-418 Sulfate is a class of aminoglycoside antibiotics, whose structure is similar to neomycin kanamycin. It can interfere with the function of 80S ribosomes in eukaryotic cells, thereby blocking protein synthesis and leading to eukaryotic cell death[1,4]. Geneticin, G-418 Sulfate from 1 to 300?μg/ml is often used for clonal selection in prokaryotes and eukaryotes. Antibiotic resistance assay of clinically isolated bacterial strains found that all strains carrying 3' -o-aminoglycoside phosphotransferases were tolerant to Geneticin, G-418 Sulfate [2].
Treatment with 100 μg/mL Geneticin, G-418 Sulfate for 12 days did not lead to the death of all SSFs, but still 44.7% of SSFs survived. Continuous treatment with 200 μg/mL Geneticin, G-418 Sulfate for 12 days can lead to the death of all SSFs. Therefore, the minimum lethal concentration of Geneticin, G-418 Sulfate leading to the death of SSFs is 200 μg/mL[5]. Geneticin, G-418 Sulfate has antiviral activity against bovine viral diarrhea virus (BVDV). Geneticin, G-418 Sulfate can prevent cytopaplasia effect (CPE) caused by DENV-2 infection of BHK cells in a dose-dependent manner, with an EC50 value of 3μg/ml[3].
In vivo, Geneticin, G-418 Sulfate destabilizes mRNAs broadly, in that the majority of mRNAs in mESCs have reduced stability when mESCs are treated with Geneticin, G-418 Sulfate. The mRNAs with half-lives that are most reduced by treatment with Geneticin, G-418 Sulfate are enriched for select optimal codons, containing G/C at the wobble position[7].The expression of antioxidant stress kinase-related genes and apoptotic genes in donor cells treated with different concentrations of Geneticin, G-418 Sulfate significantly changed, but the DNA methylation level did not change.The in vitro development efficiency of nuclear transfer embryos from Geneticin, G-418 Sulfate-treated donor cells was significantly lower than that of controls[6].
Geneticin，G-418硫酸盐是一类氨基糖苷类抗生素，其结构与新霉素卡那霉素相似。它可以干扰真核细胞中的80S核糖体功能，从而阻止蛋白质合成并导致真核细胞死亡[1,4]。通常使用浓度为1至300μg/ml的Geneticin，G-418硫酸盐进行原核和真核克隆选择。临床分离的细菌株抗生素耐药性试验发现所有携带3'-O-氨基糖苷磷酸转移酶的菌株都对Geneticin，G-418硫酸盐具有耐受性[2]。
用100μg/mL的Geneticin，G-418硫酸盐处理12天并没有导致所有SSFs死亡，但仍有44.7%的SSFs存活。连续使用200μg/mL的Geneticin，G-418硫酸盐处理12天可以导致所有SSFs死亡。因此，导致SSFs死亡的最小致命浓度为200μg/mL[5]。 Geneticin，G-418硫酸盐对牛病毒性腹泻病毒（BVDV）具有抗病毒活性。 Geneticin，G-418硫酸盐可以以剂量依赖方式防止DENV-2感染BHK细胞引起的细胞质变性效应（CPE），EC50值为3μg/ml[3]。
在体内，Geneticin、G-418硫酸盐广泛破坏mRNA的稳定性，当mESCs接受Geneticin、G-418硫酸盐处理时，大多数mRNAs的稳定性都会降低。那些半衰期最受Geneticin、G-418硫酸盐处理影响的mRNAs富含选择性优化密码子，在wobble位点包含G/C[7]。经过不同浓度Geneticin、G-418硫酸盐处理后，供体细胞中与抗氧化应激激酶相关基因和凋亡基因表达显著改变，但DNA甲基化水平没有变化。从接受了Geneticin、G-418硫酸盐处理的供体细胞克隆出来的核移植胚胎在体外发育效率明显低于对照组[6]。
References:
[1]: Bar-Nun S, Shneyour Y, et,al. G-418, an elongation inhibitor of 80 S ribosomes. Biochim Biophys Acta. 1983 Oct 13;741(1):123-7. doi: 10.1016/0167-4781(83)90018-0. PMID: 6193810.
[2]: Davies J, Jimenez A. A new selective agent for eukaryotic cloning vectors. Am J Trop Med Hyg. 1980 Sep;29(5 Suppl):1089-92. doi: 10.4269/ajtmh.1980.29.1089. PMID: 7001938.
[3]: Zhang XG, Mason PW, et,al. Antiviral activity of geneticin against dengue virus. Antiviral Res. 2009 Jul;83(1):21-7. doi: 10.1016/j.antiviral.2009.02.204. Epub 2009 Mar 11. PMID: 19501253; PMCID: PMC2694137.
[4]: Chen B., Shi X.Y., et,al. 2012, Cytoplasm vacuolization of fibroblasts during purification of Schwann cells by geneticin (G418): An optical microscope observation and analysis, Zhongguo Zhuzhi Gongcheng Yanjiu (Journal of Clinical Rehabilitative Tissue Engineering Research), 16 (14): 2593-2596
[5]: Wu Xian, JING Qian-ge, et,al. Study on the resistance of sheep skin fibroblasts to G418 and Blasticidin S [J].Genomics and applied biology,2019,38(03):1006-1011.
[6]: Li Jia-qi, Li Zi-Cong, et,al. Effects of G418 treatment on development efficiency of porcine cloned embryos in vitro [J]. Journal of south China agricultural university,2016,37(05):13-18.
[7]: Durmaz YT, Shatadal A, et,al. Geneticin reduces mRNA stability. PLoS One. 2022 Jul 28;17(7):e0272058. doi: 10.1371/journal.pone.0272058. PMID: 35901009; PMCID: PMC9333311.
| Cas No. | 108321-42-2 | SDF | |
| 别名 | G-418硫酸盐,Antibiotic G418 | ||
| 化学名 | 2-[4,6-diamino-3-[3-amino-4,5-dihydroxy-6-(1-hydroxyethyl)oxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-5-methyl-4-(methylamino)oxane-3,5-diol;sulfuric acid | ||
| Canonical SMILES | CC(C1C(C(C(C(O1)OC2C(CC(C(C2O)OC3C(C(C(CO3)(C)O)NC)O)N)N)N)O)O)O.OS(=O)(=O)O.OS(=O)(=O)O | ||
| 分子式 | C20H40N4O10.2H2SO4 | 分子量 | 692.71 |
| 溶解度 | ≥ 240.4 mg/ml in Water | 储存条件 | Store at -20°C, stored under nitrogen |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.4436 mL | 7.218 mL | 14.4361 mL |
| 5 mM | 0.2887 mL | 1.4436 mL | 2.8872 mL |
| 10 mM | 0.1444 mL | 0.7218 mL | 1.4436 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Colonic organoids derived from human induced pluripotent stem cells for modeling colorectal cancer and drug testing
Nat Med2017 Jul;23(7):878-884.PMID: 28628110DOI: 10.1038/nm.4355
With the goal of modeling human disease of the large intestine, we sought to develop an effective protocol for deriving colonic organoids (COs) from differentiated human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs). Extensive gene and immunohistochemical profiling confirmed that the derived COs represent colon rather than small intestine, containing stem cells, transit-amplifying cells, and the expected spectrum of differentiated cells, including goblet and endocrine cells. We applied this strategy to iPSCs derived from patients with familial adenomatous polyposis (FAP-iPSCs) harboring germline mutations in the WNT-signaling-pathway-regulator gene encoding APC, and we generated COs that exhibit enhanced WNT activity and increased epithelial cell proliferation, which we used as a platform for drug testing. Two potential compounds, XAV939 and rapamycin, decreased proliferation in FAP-COs, but also affected cell proliferation in wild-type COs, which thus limits their therapeutic application. By contrast, we found that Geneticin, a ribosome-binding antibiotic with translational 'read-through' activity, efficiently targeted abnormal WNT activity and restored normal proliferation specifically in APC-mutant FAP-COs. These studies provide an efficient strategy for deriving human COs, which can be used in disease modeling and drug discovery for colorectal disease.
Geneticin reduces mRNA stability
PLoS One2022 Jul 28;17(7):e0272058.PMID: 35901009DOI: 10.1371/journal.pone.0272058
Messenger RNA (mRNA) translation can lead to higher rates of mRNA decay, suggesting the ribosome plays a role in mRNA destruction. Furthermore, mRNA features, such as codon identities, which are directly probed by the ribosome, correlate with mRNA decay rates. Many amino acids are encoded by synonymous codons, some of which are decoded by more abundant tRNAs leading to more optimal translation and increased mRNA stability. Variable translation rates for synonymous codons can lead to ribosomal collisions as ribosomes transit regions with suboptimal codons, and ribosomal collisions can promote mRNA decay. In addition to different translation rates, the presence of certain codons can also lead to higher or lower rates of amino acid misincorporation which could potentially lead to protein misfolding if a substituted amino acid fails to make critical contacts in a structure. Here, we test whether Geneticin-G418, an aminoglycoside antibiotic known to promote amino acid misincorporation-affects mRNA stability. We observe that G418 decreases firefly luciferase mRNA stability in an in vitro translation system and also reduces mRNA stability in mouse embryonic stem cells (mESCs). G418-sensitive mRNAs are enriched for certain optimal codons that contain G or C in the wobble position, arguing that G418 blunts the stabilizing effects of codon optimality.
Antiviral activity of Geneticin against dengue virus
Antiviral Res2009 Jul;83(1):21-7.PMID: 19501253DOI: 10.1016/j.antiviral.2009.02.204
The aminoglycoside, Geneticin (G418), was recently shown to have antiviral activity against bovine viral diarrhea virus (BVDV). Since BVDV, dengue virus (DENV) and yellow fever virus (YFV) all belong to the Flaviviridae family, it seemed possible that a common step in their life cycle might be affected by this aminoglycoside. Here it is shown that Geneticin prevented the cytopathic effect (CPE) resulting from DENV-2 infection of BHK cells, in a dose-dependent manner with an 50% effective concentration (EC(50)) value of 3+/-0.4microg/ml. Geneticin had no detectable effect on CPE caused by YFV in BHK cells. Geneticin also inhibited DENV-2 viral yield with an EC(50) value of 2+/-0.1microg/ml and an EC(90) value of 20+/-2microg/ml. With a CC(50) value of 165+/-5microg/ml, the selectivity index of anti-DENV activity of Geneticin in BHK cells was established to be 66. Furthermore, 25microg/ml of Geneticin nearly completely blocked plaque formation induced by DENV-2, but not YFV. In addition, Geneticin, inhibited DENV-2 viral RNA replication and viral translation. Gentamicin, kanamycin, and the guanidinylated Geneticin showed no anti-DENV activity. Neomycin and paromomycin demonstrated weak antiviral activity at high concentrations. Finally, aminoglycoside-3'-phosphotransferase activity of neomycin-resistant gene abolished antiviral activity of Geneticin.