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长核酸转染试剂EntransterTM-H4000

货号规格单价数量
4000-50.75ml¥999.00

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4000-61.5ml¥1,799.00

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订单满1000元，免运费

产品介绍

EntransterTM-H4000是英格恩生物公司研发合成的纳米聚合物转染试剂，该试剂采用纳米技术合成，是最新一代非病毒转染试剂。由于纳米技术的应用，EntransterTM-H4000在细胞转染过程中，表现了卓越的低毒、高效的性能。

本品推荐用于常规DNA以及长片段（>1kb）的mRNA等RNA的转染。

推荐应用范围

常规DNA以及长片段（>1kb）的mRNA等RNA的转染。

成功转染的细胞

人源，遗传变异细胞和正常组织来源细胞 人源，肿瘤细胞 其他动物来源细胞
AsPC-1，BNL CL2，HEK293，CFPEo，HUVEC，HLMEC，H-MVEC，MRC-5，HCS-2/8，HLF，D407，H9C2，SMC A549，HuH-7，HepG2，HME4，A375，Hep-2，SW480，SKOV-3，IGROV1，K562，Jurkat，MCF7，MDA-MB-231，ZR-75-1，TCA8113，Eca-109，SGC-7901，HeLa，SiHa，Ca Ski，CaCo2，SAOS-2，PC-3，SHEP B16BL6，B16F10，C2C12，C6，CHO，COS-7，CV-1，BHK-21，MDCK，PC12，BAEC，MLE-12，MLE-15，Neuro2A，PGHAM-1，RAW 264.7，EL-4，L929，LLC，3LL，NIH 3T3

各细胞系详细转染效率情况

细胞名称 来源 转染效率(%)
AsPC-1 人源，胰脏细胞 80%
BNL CL2 人源，胚胎肾细胞 70%
HEK 293 人源，胚胎肾细胞 90%
CFPEo 人源，气管上皮细胞 50%
HUVEC 人源，脐带静脉内皮状细胞 55%
HLMEC 人源，肺血管内皮细胞 60%
H-MVEC 人源，微血管内皮细胞 50%
MRC-5 人源，胚胎肺细胞 75%
HCS-2/8 人源，软骨样细胞 40%
HLF 人源，肺成纤维细胞 65%
D 407 人源，视网膜色素上皮细胞 60%
A549 人源，肺癌细胞 90%
HuH-7 人源，肝癌细胞 63%
HepG2 人源，肝癌细胞 30%
HME4 人源，黑色素瘤细胞 90%
A375 人源，黑色素瘤细胞 70%
Hep-2 人源，喉癌细胞 70%
SAOS-2 人源，骨肉瘤细胞 50%
SKOV-3 人源，卵巢腺癌细胞 60%
IGROV1 人源，卵巢癌细胞 20%
K562 人源，慢性白血病细胞 30%
Jurkat 人源，T淋巴细胞白血病细胞 45%
MCF7 人源，乳腺癌细胞 55%
MDA-MB-231 人源，乳腺癌细胞 70%
ZR-75-1 人源，乳腺癌细胞 70%
TCA8113 人源，舌鳞癌细胞 90%
Eca-109 人源，胃腺癌细胞 57%
SGC-7901 人源，胃腺癌细胞 48%
BGC-823 人源，胃腺癌细胞 60%
Panc-1 人源，胰腺癌细胞 40%
HeLa 人源，宫颈癌细胞 80%
SiHa 人源，宫颈癌细胞 70%
Ca Ski 人源，宫颈表皮样癌细胞 80%
CaCo2 人源，结肠癌上皮细胞 30%
SW480 人源，结肠腺癌细胞 80%
PC-3 人源，前列腺癌细胞 30%
SHEP 人源，神经母细胞瘤细胞 70%
B16BL6 小鼠，黑色素瘤细胞 90%
B16F10 小鼠，黑色素瘤细胞 80%
C2C12 小鼠，胚胎肌原细胞 30%
C6 大鼠，脑胶质瘤细胞 60%
CHO 中国仓鼠，卵巢细胞 90%
COS-7 猴SV40转化肾细胞 80%
CV-1 非洲绿猴肾细胞 70%
BHK-21 仓鼠，肾细胞 80%
MDCK 犬，肾上皮细胞 30%
PC12 大鼠，肾上腺嗜铬细胞瘤细胞 33%
BAEC 牛，动脉内皮细胞 50%
MLE-12 小鼠，肺上皮细胞 70%
MLE-15 小鼠，肺上皮细胞 70%
Neuro2A 小鼠，成神经瘤细胞 70%
PGHAM-1 仓鼠，胰腺癌细胞 90%
RAW 264.7 小鼠，巨噬细胞 15%
EL-4 小鼠，胸腺瘤细胞 75%
L929 小鼠，成纤维肉瘤细胞 60%
LLC 小鼠，路易斯肺癌细胞 75%
3LL 小鼠，路易斯肺癌细胞 65%
NIH 3T3 小鼠，胚胎纤维原细胞 40%
P19 小鼠，胚胎肉瘤细胞 60%

主要特点

用法用量和常见的脂质体试剂完全一样，可不改流程方法更换试剂 转染效率更高，每支节省1000元以上，可根据需要定做500ml以上大包装，可批量定制产品。

  1. Cell Death Dis. 2026 May 16.doi: 10.1038/s41419-026-08872-1. Targeting lysine-specific demethylase 1 inhibits melanoma metastasis via the NF2-Hippo-YAP pathway 靶向赖氨酸特异性脱甲基酶1可通过NF2-Hippo-YAP通路抑制黑色素瘤转移 (IF:9.6).

  2. Nat Commun. 2026 Apr 20.doi: 10.1038/s41467-026-72181-6. Photoactivatable CRISPR/Cas13d via upconversion nanoparticles for deep tissue RNA engineering and orthopedic therapy 通过上转换纳米颗粒实现光激活CRISPR/Cas13d，用于深层组织RNA工程和骨科治疗(IF:15.7).

  3. Viruses. 2026 Mar 5;18(3):323.doi: 10.3390/v18030323. Tree Shrew Genome-Wide CRISPR Screen Identifies RNF6 as a Proviral Host Factor for Zika Virus Replication in Brain Microvascular Endothelial Cells 树鼩全基因组CRISPR筛选鉴定出RNF6是寨卡病毒在脑微血管内皮细胞中复制的病毒前体宿主因子

  4. Cell Rep Methods. 2026 Feb 23;6(2):101299.doi: 10.1016/j.crmeth.2025.101299. Epub 2026 Feb 11. An orthogonal CRISPR/Cpf1 platform for precise spatiotemporal gene regulation and osteoporotic fracture repair 用于精确时空基因调控和骨质疏松性骨折修复的正交CRISPR/Cpf1平台 (IF:4.500).

  5. Biochem Pharmacol. 2025 Dec;242(Pt 4):117403.doi: 10.1016/j.bcp.2025.117403. Epub 2025 Oct 3. Plectin promotes bone formation via phase separation and sequestering annexin A2 Plec通过相分离和螯合Anxa2来促进骨形成 (IF:5.3).

  6. Aging Cell. 2025 Dec 9:e70306.doi: 10.1111/acel.70306. FoxO1 Responses to Chronic Oxidative Stress to Participate in Age-Related Osteoporosis by Depriving β-Catenin From TCF7 。FoxO1对慢性氧化应激的反应通过从TCF7中剥夺β-Catenin参与年龄相关性骨质疏松症 (IF:8.104).

  7. Biochem Pharmacol. 2025 Oct 3;242(Pt 4):117403.doi: 10.1016/j.bcp.2025.117403 Plectin promotes bone formation via phase separation and sequestering annexin A2 Plec通过相分离和螯合Anxa2促进骨形成(IF:5.6).

  8. Gene. 2025 Sep 15:965:149650.doi: 10.1016/j.gene.2025.149650. Epub 2025 Jun 30. Contribution of cysteamine dioxygenase to taurine biosynthesis in the oyster Crassostrea gigas半胱胺双加氧酶对牡蛎牛磺酸生物合成的贡献

  9. Trends Biotechnol. 2025 Aug 30:S0167-7799(25)00314-2.doi: 10.1016/j.tibtech.2025.07.029. Highly efficient prime editors for mammalian genome editing based on porcine retrovirus reverse transcriptase 基于猪逆转录病毒逆转录酶的高效哺乳动物基因组编辑引物 (IF:14.900).

  10. Comp Biochem Physiol B Biochem Mol Biol. 2025 Aug-Sep:279:111128.doi: 10.1016/j.cbpb.2025.111128. Epub 2025 Jul 10.
    Metallothionein CgMTIII is involved in zinc binding and accumulation in the Pacific oyster Crassostrea gigas 金属硫蛋白CgMTIII参与太平洋牡蛎Crassostrea gigas中锌的结合和积累

  11. Adv Sci (Weinh). 2025 Jul 14:e06492.doi: 10.1002/advs.202506492. Genomic Insights into the Origin, High Fecundity and Environmental Adaptation of Hu Sheep 湖羊起源、高繁殖力和环境适应性的基因组学研究 (IF:15.1).

  12. Sci Rep. 2025 Jul 1;15(1):20687.doi: 10.1038/s41598-025-05826-z. LncRNA CTD-2555A7.2 promotes bone formation with LncRNA-specific cascade amplification strategy LncRNA CTD-2555A7.2通过LncRNA特异性级联扩增策略促进骨形成

  13. J Ethnopharmacol. 2025 Jun 12:349:119935.doi: 10.1016/j.jep.2025.119935. Epub 2025 May 7. Mechanism of Rabdosia rubescens extract against gastric cancer microenvironment by SIRT1/NF-κB/p53 pathway and promoting tumor-associated macrophage polarization 冬凌草提取物通过SIRT1/NF-κB/p53途径抑制癌症微环境及促进肿瘤相关巨噬细胞极化的机制

  14. J Invertebr Pathol. 2025 Jun:210:108289.doi: 10.1016/j.jip.2025.108289. Epub 2025 Feb 21. Characterization and functional analysis of the small heat shock protein HSP19.5 in Bombyx mori in response to Nosema bombycis infection 小分子热休克蛋白HSP19的特性和功能分析。家蚕对家蚕微孢子虫感染的反应

  15. Commun Biol . 2025 May 29;8(1):827.doi: 10.1038/s42003-025-08290-7. A new type of Caspase-1 upon recognizing bacteria inhibits GSDME-dependent histone modification and NF-κB signaling 一种新型的Caspase-1在识别细菌时抑制GSDME依赖的组蛋白修饰和NF-κB信号传导 (IF:5.2).

  16. Insects. 2025 May 21;16(5):544.doi: 10.3390/insects16050544. 20-Hydroxyecdysone Modulates Bmp53-Mediated Apoptosis Regulation by Suppressing Mdm2-like-Dependent Ubiquitination in Silkworm, Bombyx mori

  17. Biotechnol Lett. 2025 Apr 16;47(3):42.doi: 10.1007/s10529-025-03576-6. Construction and immunogenicity analysis of a recombinant baculovirus targeting the N protein of SARS-CoV-2 靶向SARS-CoV-2 N蛋白的重组杆状病毒的构建及免疫原性分析

  18. Phytomedicine. 2025 Apr:139:156442.doi: 10.1016/j.phymed.2025.156442. Epub 2025 Feb 1. Naturally-occurring carnosic acid as a promising therapeutic agent for skin inflammation via targeting STAT1 天然鼠尾草酸作为靶向STAT1治疗皮肤炎症的有前景的药物 (IF:6.7).

  19. PLoS Biol. 2025 Apr 1;23(4):e3003047.doi: 10.1371/journal.pbio.3003047. eCollection 2025 Apr. Mdga2 deficiency leads to an aberrant activation of BDNF/TrkB signaling that underlies autism-relevant synaptic and behavioral changes in mice Mdga2缺乏导致BDNF/TrkB信号的异常激活，这是小鼠自闭症相关突触和行为变化的基础(IF:9.8).

  20. Bull Entomol Res. 2025 Mar 24:1-14.doi: 10.1017/S000748532500015X. BmWARS inhibits BmNPV infection via the PI3K-Akt pathway BmWARS通过PI3K-Akt途径抑制BmNPV感染

  21. J Invertebr Pathol. 2025 Feb 21:210:108289.doi: 10.1016/j.jip.2025.108289. Characterization and functional analysis of the small heat shock protein HSP19.5 in Bombyx mori in response to Nosema bombycis infection 小分子热休克蛋白HSP19.5的特性和功能分析对家蚕微孢子虫感染的反应

  22. Cancer Cell Int. 2025 Jan 13;25(1):10.doi: 10.1186/s12935-025-03638-9.Heterogeneous nuclear ribonucleoprotein C promotes non-small cell lung cancer progression by enhancing XB130 mRNA stability and translation (IF:5.3).异质性核核糖核蛋白C通过增强XB130mRNA的稳定性和翻译促进非小细胞肺癌的进展

  23. J Virol. 2024 Nov 27:e0151124.doi: 10.1128/jvi.01511-24. Unraveling dual fusion mechanisms in BmNPV GP64: critical roles of CARC motifs and signal peptide retention 解开BmNPV GP64中的双重融合机制：CARC基序和信号肽保留的关键作用

  24. Mar Biotechnol (NY). 2024 Nov 27;27(1):11.doi: 10.1007/s10126-024-10379-9. A Zinc Uptake Transporter ZIP1-II Is Involved in Zinc Accumulation in the Hepatopancreas of Pacific Oyster Crassostrea gigas 锌摄取转运蛋白ZIP1-II参与太平洋牡蛎肝胰腺中锌的积累

  25. Microb Cell Fact. 2024 Oct 18;23(1):284.doi: 10.1186/s12934-024-02534-7. The signal peptide of BmNPV GP64 activates the ERAD pathway to regulate heterogeneous secretory protein expression BmNPV GP64信号肽激活ERAD通路调节异质性分泌蛋白表达

  26. Virology. 2024 Sep:597:110147.doi: 10.1016/j.virol.2024.110147. Epub 2024 Jun 19. Unveiling non-classical glycosylation patterns in Bombyx mori nucleopolyhedrovirus GP64: Insights into viral entry and fusion 揭示家蚕核型多角体病毒GP64的非经典糖基化模式：对病毒进入和融合的见解

  27. Front Pharmacol. 2024 Sep 6:15:1439497.doi: 10.3389/fphar.2024.1439497. eCollection 2024. The lncRNA CADM2-AS1 promotes gastric cancer metastasis by binding with miR-5047 and activating NOTCH4 translation lncRNA CADM2-AS1通过与miR-5047结合并激活NOTCH4翻译来促进癌症转移 (IF:5.6).

  28. Biol Res. 2024 Sep 9;57(1):64.doi: 10.1186/s40659-024-00544-8. Mouse testicular macrophages can independently produce testosterone and are regulated by Cebpb 小鼠睾丸巨噬细胞可以独立产生睾酮，并受Cebpb的调节

  29. J Virol Methods. 2024 Jun:327:114933.doi: 10.1016/j.jviromet.2024.114933. Epub 2024 Apr 4. Intracellular localized heterogeneous protein franking by a transmembrane domain of GP64 is sufficient to be assembled on budded virions of Bombyx mori nucleopolyhedrovirus GP64跨膜结构域在细胞内定位的异质蛋白franking足以在家蚕核型多角体病毒的芽状病毒颗粒上组装

  30. Int J Biol Macromol. 2024 May;266(Pt 1):131197.doi: 10.1016/j.ijbiomac.2024.131197. Epub 2024 Mar 28. Bombyx mori triose-phosphate transporter protein inhibits Bombyx mori nucleopolyhedrovirus infection by reducing the cell glycolysis pathway 家蚕三糖磷酸转运蛋白通过减少细胞糖酵解途径抑制家蚕核型多角体病毒感染

  31. Invest Ophthalmol Vis Sci. 2024 Apr 1;65(4):1.doi: 10.1167/iovs.65.4.1. Identification of Novel FZD4 Mutations in Familial Exudative Vitreoretinopathy and Investigating the Pathogenic Mechanisms of FZD4 Mutations 家族性渗出性玻璃体视网膜病中新型FZD4突变的鉴定及FZD4突变致病机制的研究

  32. Int Immunopharmacol. 2024 Apr 20:131:111864.doi: 10.1016/j.intimp.2024.111864. Epub 2024 Mar 13. miR-186-5p improves alveolar epithelial barrier function by targeting the wnt5a/β-catenin signaling pathway in sepsis-acute lung injury miR-186-5p通过靶向wnt5a/β-catenin信号通路改善脓毒症急性肺损伤肺泡上皮屏障功能

  33. J Exp Clin Cancer Res. 2024 Apr 23;43(1):123.doi: 10.1186/s13046-024-03048-1. Tumor-suppressive miR-4732-3p is sorted into fucosylated exosome by hnRNPK to avoid the inhibition of lung cancer progression (IF:12.658).肿瘤抑制性miR-4732-3p通过hnRNPK分类为岩藻糖基外泌体，以避免对肺癌癌症进展的抑制

  34. Mol Genet Genomics. 2024 Mar 13;299(1):32.doi: 10.1007/s00438-024-02128-3. Characterization of a novel heterozygous frameshift variant in NDP gene that causes familial exudative vitreoretinopathy in female patients 导致女性家族性渗出性玻璃体视网膜病变的NDP基因新型杂合移码变异的特征

  35. Invest Ophthalmol Vis Sci. 2024 Mar 5;65(3):31.doi: 10.1167/iovs.65.3.31. Investigating the Impact of Dimer Interface Mutations on Norrin’s Secretion and Norrin/β-Catenin Pathway Activation 二聚体界面突变对Norrin分泌和Norrin/β-Catenin通路激活的影响

  36. Dev Comp Immunol. 2024 Mar:152:105114.doi: 10.1016/j.dci.2023.105114. Epub 2023 Dec 13. Ras3 in Bombyx mori with antiviral function against B. mori nucleopolyhedrovirus 家蚕Ras3对家蚕核型多角体病毒具有抗病毒作用

  37. FASEB J. 2024 Feb 29;38(4):e23493.doi: 10.1096/fj.202302387R. Deciphering a crucial dimeric interface governing Norrin dimerization and the pathogenesis of familial exudative vitreoretinopathy 解读Norrin二聚化的关键二聚体界面和家族性渗出性玻璃体视网膜病变的发病机制

  38. Int J Biol Macromol. 2024 Feb;258(Pt 1):128570.doi: 10.1016/j.ijbiomac.2023.128570. Epub 2023 Dec 12. Frameshift variants in the C-terminal of CTNNB1 cause familial exudative vitreoretinopathy by AXIN1-mediated ubiquitin-proteasome degradation condensation (IF:8.2. CTNNB1 C末端的移码变异通过AXIN1介导的泛素-蛋白酶体降解缩合引起家族性渗出性玻璃体视网膜病变

  39. Cell Commun Signal. 2024 Feb 1;22(1):93.doi: 10.1186/s12964-024-01497-x. Mechanosensitive channel of large conductance enhances the mechanical stretching-induced upregulation of glycolysis and oxidative metabolism in Schwann cells (IF:7.525). 大电导的机械敏感通道增强了机械拉伸诱导的雪旺氏细胞糖酵解和氧化代谢的上调

  40. Small Methods. 2024 Jan 2:e2301310.doi: 10.1002/smtd.202301310. Online ahead of print. An Inducible CRISPR-dCas9-Based Transcriptional Repression System for Cancer Therapy 一种用于癌症治疗的基于诱导CRISPR-dCas9的转录抑制系统(IF:25.367).

  41. Cell Death Dis. 2023 Nov 3;14(11):716.doi: 10.1038/s41419-023-06238-5. Inhibition of LSD1 induces ferroptosis through the ATF4-xCT pathway and shows enhanced anti-tumor effects with ferroptosis inducers in NSCLC CTNNB1新截短变异体引起家族性渗出性玻璃体视网膜病变

  42. Mol Neurobiol. 2023 Nov 22.doi: 10.1007/s12035-023-03808-8. Mol Neurobiol . 2023 Nov 22. doi: 10.1007/s12035-023-03808-8. Online ahead of print. GSK-126 Attenuates Cell Apoptosis in Ischemic Brain Injury by Modulating the EZH2-H3K27me3-Bcl2l1 Axis GSK-126通过调节EZH2-H3K27me3-Bcl2l1轴减轻缺血性脑损伤中的细胞凋亡

  43. Int J Biol Macromol. 2023 Sep 7:126801.doi: 10.1016/j.ijbiomac.2023.126801. Online ahead of print. Neddylation-dependent Neddylation-dependent LSD1 destabilization inhibits the stemness and chemoresistance of gastric cancer LSD1失稳抑制癌症的干燥和化疗耐药性

  44. Gene. 2023 Sep 25:881:147626.doi: 10.1016/j.gene.2023.147626. Epub 2023 Jul 8. BmINR and BmAC6 genes involve in diapause regulation via the insulin/IGF signaling pathway in the silkworm (Bombyx mori) 家蚕BmINR和BmAC6基因通过胰岛素/IGF信号通路参与滞育调控

  45. Sci Adv. 2023 Jul 21;9(29):eadf7858.doi: 10.1126/sciadv.adf7858. Epub 2023 Jul 21. Engineered exosomes reprogram Gli1+ cells in vivo to prevent calcification of vascular grafts and autologous pathological vessels 工程外泌体在体内重新编程Gli1+细胞，以防止血管移植物和自体病理血管钙化 (IF:14.98).

  46. Mol Biol Rep. 2023 Jun;50(6):5295-5306.doi: 10.1007/s11033-023-08489-z. Epub 2023 May 6. m6A-dependent mevalonate kinase in juvenile hormone synthesis pathway regulates the diapause process of bivoltine silkworm (Bombyx mori) m6A依赖性甲羟戊酸激酶在幼激素合成途径中调节二伏蚕滞育过程

  47. Food Sci Biotechnol. 2023 Jun 19;33(2):465-474.doi: 10.1007/s10068-023-01358-2. eCollection 2024 Jan. A novel protein extracted from Hemerocallis citrina Borani inhibits hepatocellular carcinoma cell proliferation by regulating mitochondria-dependent apoptosis and aerobic glycolysis 从萱草中提取的一种新蛋白通过调节线粒体依赖性细胞凋亡和抑制肝癌细胞增殖

  48. Insects. 2023 Apr 5;14(4):362.doi: 10.3390/insects14040362. Determination of Key Components in the Bombyx mori p53 Apoptosis Regulation Network Using Y2H-Seq Y2H-Seq法测定家蚕p53细胞凋亡调控网络的关键成分

  49. Oncol Lett. 2023 Feb 28;25(4):143.doi: 10.3892/ol.2023.13729. eCollection 2023 Apr. Effects of Helicobacter pylori on the expression of the FTO gene and its biological role in gastric cancer 幽门螺杆菌对FTO基因表达的影响及其在癌症中的生物学作用

  50. J Med Genet. 2023 Feb;60(2):174-182.doi: 10.1136/jmedgenet-2021-108259. Epub 2022 Mar 31. Novel truncating variants in CTNNB1 cause familial exudative vitreoretinopathy CTNNB1新截短变异体引起家族性渗出性玻璃体视网膜病变

  51. Biochem Biophys Res Commun. 2023 Jan 8:639:36-45.doi: 10.1016/j.bbrc.2022.11.067. Epub 2022 Nov 24. Silencing long non-coding RNA SNHG3 repairs the dysfunction of pulmonary microvascular endothelial barrier by regulating miR-186-5p/Wnt axis 沉默长非编码RNA SNHG3通过调节miR-186-5p/Wnt轴修复肺微血管内皮屏障功能障碍

  52. Arch Pharm Res. 2023 Jan;46(1):44-58.doi: 10.1007/s12272-023-01426-5. Epub 2023 Jan 6. MEKs/ERKs-mediated FBXO1/E2Fs interaction interference modulates G1/S cell cycle transition and cancer cell proliferation MEKs/ERKs介导的FBXO1/E2Fs相互作用干扰调节G1/S细胞周期转变和癌症细胞增殖

  53. Front Surg. 2023 Jan 6:9:1062659.doi: 10.3389/fsurg.2022.1062659. eCollection 2022. Classification of the mitochondrial ribosomal protein-associated molecular subtypes and identified a serological diagnostic biomarker in hepatocellular carcinoma 肝细胞癌线粒体核糖体蛋白相关分子亚型的分类及血清学诊断标志物的鉴定

  54. Adipocyte. 2022 Dec;11(1):477-486.doi: 10.1080/21623945.2022.2107786. (IF3.553) MAPKs/AP-1, not NF-κB, is responsible for MCP-1 production in TNF-α-activated adipocytes 激活的脂肪细胞中MCP-1的产生

  55. Int Immunopharmacol. 2022 Dec;113(Pt A):109312.doi: 10.1016/j.intimp.2022.109312. Epub 2022 Oct 14. Resibufogenin, one of bufadienolides in toad venom, suppresses LPS-induced inflammation via inhibiting NF-κB and AP-1 pathways CDK1通过抑制∆Np63α介导的转录调控促进头颈部鳞状细胞癌细胞的上皮-间充质过渡和迁移

  56. Protein Expr Purif. 2022 Dec:200:106156.doi: 10.1016/j.pep.2022.106156. Epub 2022 Aug 18. The cytoplasmic tail substitution increases the assembly efficiency of Ebola virus glycoprotein on the budded virus of Bombyx mori nucleopolyhedrovirus 细胞质尾部置换提高了埃博拉病毒糖蛋白在家蚕核型多角体病毒芽状病毒上的组装效率

  57. Front Mol Neurosci. 2022 Dec 8:15:1034766.doi: 10.3389/fnmol.2022.1034766. eCollection 2022. Nucleosome assembly protein 1-like 5 alleviates Alzheimer’s disease-like pathological characteristics in a cell model 核小体组装蛋白1-like 5在细胞模型中缓解阿尔茨海默病样病理特征

  58. Microbiol Spectr. 2022 Aug 31;10(4):e0191322.doi: 10.1128/spectrum.01913-22. Epub 2022 Aug 8. (IF9.043) The Bombyx mori Nucleopolyhedrovirus GP64 Retains the Transmembrane Helix of Signal Peptide to Contribute to Secretion across the Cytomembrane 家蚕核多角体病毒GP64保留信号肽的跨膜螺旋以促进跨细胞膜分泌

  59. Fish Shellfish Immunol. 2022 Aug:127:129-139.doi: 10.1016/j.fsi.2022.05.057. Epub 2022 Jun 14. A RAC-alpha serine/threonine-protein kinase (CgAKT1) involved in the synthesis of CgIFNLP in oyster Crassostrea gigas RACα-丝氨酸/苏氨酸蛋白激酶（CgAKT1）参与牡蛎CgIFNLP的合成

  60. Research (Wash D C) . 2022 Jul 19;2022:9826426. doi: 10.34133/2022/9826426. eCollection 2022. Programming of Regulatory T Cells In Situ for Nerve Regeneration and Long-Term Patency of Vascular Grafts. (IF:11.036).

  61. Int J Mol Sci. 2022 Jul 2;23(13):7385.doi: 10.3390/ijms23137385. (IF6.208) CDK1 Promotes Epithelial-Mesenchymal Transition and Migration of Head and Neck Squamous Carcinoma Cells by Repressing ∆Np63α-Mediated Transcriptional Regulation CDK1通过抑制∆Np63α介导的转录调节促进头颈部鳞状细胞的上皮-间充质转移和迁移

  62. J Med Genet. 2022 Mar 31;jmedgenet-2021-108259. doi: 10.1136/jmedgenet-2021-108259. Online ahead of print. Novel truncating variants in CTNNB1 cause familial exudative vitreoretinopathy (IF:6.318).

  63. STAR Protoc. 2022 Mar 15;3(2):101239.doi: 10.1016/j.xpro.2022.101239. eCollection 2022 Jun 17. Measuring Laplace pressure and imaging the actin cortex during cytokinesis in cultured cells 培养细胞胞质分裂过程中拉普拉斯压力的测量和肌动蛋白皮层的成像

  64. Food Funct. 2022 Mar 7;13(5):2913-2924. doi: 10.1039/d1fo02755g. and bone formation via activating the Wnt/β-catenin signaling pathway (IF:5.396).

  65. Ticks Tick Borne Dis. 2022 Jan 17;13(3):101906.Differential characteristics of mammalian and tick-derived promoters to trigger protein expression in transfected tick cell lines.

  66. Dev Comp Immunol. 2022 Jun;131:104382. doi: 10.1016/j.dci.2022.104382. Epub 2022 Mar 1. A cypovirus encoded microRNA negatively regulates the NF-κB pathway to enhance viral multiplication in Silkworm, Bombyx mori

  67. Exp Mol Med.2022 Jan;54(1):35-46.doi: 10.1038/s12276-021-00721-9.Epub 2022 Jan 12. FBXW7-mediated ERK3 degradation regulates the proliferation of lung cancer cells.

  68. Nutr Cancer. 2022;74(1):333-345.doi: 10.1080/01635581.2021.1882508. Epub 2021 Feb 5. TKP, a Serine Protease from Trichosanthes kirilowii, Inhibits Cell Proliferation by Blocking Aerobic Glycolysis in Hepatocellular Carcinoma Cells 天花粉丝氨酸蛋白酶TKP通过阻断肝细胞癌细胞的有氧糖酵解抑制细胞增殖

  69. iScience. 2022 Feb 21;25(3):103949. doi: 10.1016/j.isci.2022.103949. eCollection 2022 Mar 18. Long noncoding RNA Lnc-DIF inhibits bone formation by sequestering miR-489-3p (IF:6.007).

  70. Eur J Ophthalmol. 2022 Jan 17;11206721221074209. doi: 10.1177/11206721221074209. Online ahead of print. Whole-Exome Sequencing Reveals Novel NDP Variants in X-Linked Familial Exudative Vitreoretinopathy

  71. J Anim Sci Biotechnol. 2021 Dec 13;12(1):120. doi: 10.1186/s40104-021-00639-2. Transcriptome sequencing analysis for the identification of stable lncRNAs associated with bovine Staphylococcus aureus mastitis (IF:5.032).

  72. Arch Pharm Res. 2021 Dec;44(12):1091-1108.doi: 10.1007/s12272-021-01358-y. Epub 2021 Nov 9. Kaempferol sensitizes cell proliferation inhibition in oxaliplatin-resistant colon cancer cells 山奈酚对奥沙利铂耐药结肠癌癌症细胞增殖抑制的敏感性

  73. Insect Mol Biol. 2021 Dec;30(6):552-565.doi: 10.1111/imb.12725. Epub 2021 Jul 30. Functional analysis of a putative Bombyx mori cypovirus miRNA BmCPV-miR-10 and its effect on virus replication 家蚕cypovirus miRNA BmCPV-miR-10的功能分析及其对病毒复制的影响

  74. Biomed Pharmacother. 2021 Oct;142:112056.doi:10.1016/j.biopha.2021.112056. Epub 2021 Aug 23. Dysregulation of miR-342-3p in plasma exosomes derived from convalescent AMI patients and its consequences on cardiac repair恢复期AMI患者血浆外小体中miR-342-3p的失调及其对心脏修复的影响

  75. Fish Shellfish Immunol. 2021 Dec;119:173-181. doi: 10.1016/j.fsi.2021.09.016. Epub 2021 Oct 2. Identification and characterization of an apoptosis-inducing factor 1 involved in apoptosis and immune defense of oyster, Crassostrea gigas

  76. Photodermatol Photoimmunol Photomed. 2021 Sep;37(5):395-403.doi: 10.1111/phpp.12670. Epub 2021 Feb 23. Ultraviolet B irradiation up-regulates MM1 and induces photoageing of the epidermis 紫外线B照射上调MM1并诱导表皮光老化

  77. Anal Chem. 2021 Aug 31;93(34):11775-11784.doi: 10.1021/acs.analchem.1c02197. Epub 2021 Aug 19. Facile Method for Specifically Sensing Sphingomyelinase in Cells and Human Urine Based on a Ratiometric Fluorescent Nanoliposome Probe 基于定量荧光纳米脂质体探针的简便特异性检测细胞和人尿液中鞘氨醇酶的方法

  78. Cell Biosci. 2021 Aug 4;11(1):156.doi: 10.1186/s13578-021-00669-3. Modaline sulfate promotes Oct4 expression and maintains self-renewal and pluripotency of stem cells through JAK/STAT3 and Wnt signaling pathways通过JAKSTAT3和Wnt检测干细胞

  79. Front Physiol. 2021 Aug 4;12:663482.doi:10.3389/fphys.2021.663482. eCollection 2021. Two Putative Cypovirus-Encoded miRNAs Co-regulate the Host Gene of GTP-Binding Nuclear Protein Ran and Facilitate Virus ReplicationmiRNA共同调节宿主基因GTP结合核蛋白Ran的表达

  80. J Cell Mol Med. 2021 Jul;25(13):6242-6257.doi: 10.1111/jcmm.16579. Epub 2021 Jun 15. MACF1 alleviates aging-related osteoporosis via HES1 MACF1通过HES1缓解与衰老相关的骨质疏松症

  81. Cell Death Differ. 2021 Jul;28(7):2160-2178.doi:10.1038/s41418-021-00744-9. Epub 2021 Mar 4. MACF1 promotes osteoblast differentiation by sequestering repressors in cytoplasmMACF1通过隔离促进成骨细胞分化

  82. Genet Test Mol Biomarkers. 2021 Jun;25(6):399-404.doi: 10.1089/gtmb.2021.0019. Epub 2021 Jun 2.Whole-Exome Sequencing Reveals Novel TSPAN12 Variants in Autosomal Dominant Familial Exudative Vitreoretinopathy

  83. Front Pharmacol. 2021 Jun 14;12:696729.doi:10.3389/fphar.2021.696729. eCollection 2021. Forsythiasides-Rich Extract From Forsythiae Fructus Inhibits Mast Cell Degranulation by Enhancing Mitochondrial Ca 2+ Uptake连翘抑制肥大细胞生长线粒体钙摄取

  84. Biochem Pharmacol. 2021 Apr;186:114476.doi:10.1016/j.bcp.2021.114476. Epub 2021 Feb 17. Aloe-emodin, a naturally occurring anthraquinone, is a highly potent mast cell stabilizer through activating mitochondrial calcium uniporter芦荟大黄素

  85. Gene. 2021 Apr 20;777:145450.doi: 10.1016/j.gene.2021.145450. Epub 2021 Jan 29. A 14-amino acids deletion in BmShadow results to non-moult on the 2nd instar in the bivoltine silkworm, Bombyx moriBmShadow中14个氨基酸的缺失导致二伏家蚕2龄时不蜕皮

  86. FEBS Open Bio. 2021 Mar;11(3):890-897.doi: 10.1002/2211-5463.13109. Epub 2021 Feb 19. Pin1 and JNK1 cooperatively modulate TAp63γ Pin1和JNK1协同调节TAp63γ

  87. Curr Microbiol. 2021 Feb;78(2):490-501.doi: 10.1007/s00284-020-02309-4. Epub 2021 Jan 2. 18 Additional Amino Acids of the Signal Peptide of the Bombyx mori Nucleopolyhedrovirus GP64 Activates Immunoglobulin Binding Protein (BiP) Expression by RNA-seq Analysis RNA-seq分析家蚕核多角体病毒GP64信号肽的18个额外氨基酸激活免疫球蛋白结合蛋白（BiP）的表达

  88. ACS Appl Mater Interfaces. 2021 Feb 10;13(5):6043-6052.doi: 10.1021/acsami.0c21223. Epub 2021 Feb 1.A CRISPR-Cas9-Based Near-Infrared Upconversion-Activated DNA Methylation Editing System基于CRISPR-Cas9的近红外上转换激活DNA甲基化编辑系统 (IF:8.3).

  89. Pharmacol Res. 2021 Jan;163:105230.doi:10.1016/j.phrs.2020.105230. Epub 2020 Oct 5. Long noncoding RNA AK039312 and AK079370 inhibits bone formation via miR-199b-5p长非编码RNA AK039312和AK079370抑制骨形成

  90. Cell Death Differ. 2021 Jan;28(1):337-348.doi: 10.1038/s41418-020-00620-y. Epub 2020 Sep 9. RPS23RG1 modulates tau phosphorylation and axon outgrowth through regulating p35 proteasomal degradation RPS23RG1通过调节p35蛋白酶体降解调节tau磷酸化和轴突生长

  91. Pharmacol Res. 2020 Oct 5;105230. doi: 10.1016/j.phrs.2020.105230 Long noncoding RNA AK039312 and AK079370 inhibits bone formation via miR-199b-5p

  92. Biochem Biophys Res Commun. 2020 Dec10;533(3):613-619.doi:10.1016/j.bbrc.2020.09.063. Epub 2020 Sep 25. Prefoldin subunit MM1 promotes cell migration via facilitating filopodia formation预折叠蛋白亚单位MM1通过促进细胞迁移

  93. Graefes Arch Clin Exp Ophthalmol. 2020 Oct;258(10):2251-2261.doi: 10.1007/s00417-020-04636-5.Epub 2020 Jun 7.Disease-causing mutations associated with bestrophinopathies promote apoptosis in retinal pigment epithelium cells.视网膜色素上皮细胞凋亡

  94. Biochem Pharmacol. 2020 Jul;177:113984.doi:10.1016/j.bcp.2020.113984. Epub 2020 Apr 18. Ethyl-p-methoxycinnamate enhances oct4 expression and reinforces pluripotency through the NF-κB signaling pathway对甲氧基肉桂酸乙酯增强oct4表达并增强多能性

  95. Small.2020 Jul;16(30):e1907301.doi: 10.1002/smll.201907301. Epub 2020 Jun 25.Photoactivatable RNA N 6 -Methyladenosine Editing with CRISPR-Cas13甲基腺苷编辑(IF:13.281).

  96. Small. 2020 Jul;16(30):e1907301.doi: 10.1002/smll.201907301. Epub 2020 Jun 25. Photoactivatable RNA N6 -Methyladenosine Editing with CRISPR-Cas13 CRISPR-Cas13编辑可光激活RNA N6-甲基腺苷 (IF:13.3).

  97. Front Genet.2020 May 26;11:511.doi: 10.3389/fgene.2020.00511. eCollection 2020.MICA ∗ 012:01 Allele Facilitates the Metastasis of KRAS-Mutant Colorectal Cancer.KRAS突变型结直肠癌的转移

  98. J Ethnopharmacol. 2020 Mar 25:250:112499.doi: 10.1016/j.jep.2019.112499. Epub 2019 Dec 23. Ethanol extract of Elephantopus scaber Linn. Attenuates inflammatory response via the inhibition of NF-κB signaling by dampening p65-DNA binding activity in lipopolysaccharide-activated macrophages 非洲象的乙醇提取物。通过抑制脂多糖激活的巨噬细胞中p65 DNA结合活性抑制NF-κB信号传导来减轻炎症反应 (IF:5.195).

  99. J Cell Biochem. 2019 Sep;120(9):14326-14335.doi: 10.1002/jcb.28538. Epub 2019 May 20. Bom-miR-2805 upregulates the expression of Bombyx mori fibroin light chain gene in vivo. Bom-miR-2805上调体内家蚕丝素轻链基因的表达。

  100. Genes Dis.2019 Sep 17;8(1):94-109.doi: 10.1016/j.gendis.2019.09.005. eCollection 2021 Jan.SFs mediate the function of AR in the ovarian granulosa cells of patients with PCOS.SRSFs介导多囊卵巢综合征患者卵巢颗粒细胞AR的功能

  101. Genes Dis. 2019 Sep 17;8(1):94-109.doi: 10.1016/j.gendis.2019.09.005. eCollection 2021 Jan. SRSFs mediate the function of AR in the ovarian granulosa cells of patients with PCOS SRSFs介导多囊卵巢综合征患者卵巢颗粒细胞AR的功能 (IF:6.9).

  102. J Cell Physiol . 2019 Jul;234(7):11524-11536. doi: 10.1002/jcp.27815. Epub 2019 Jan 17. A novel long noncoding RNA AK016739 inhibits osteoblast differentiation and bone formation

  103. Eur Rev Med Pharmacol Sci.2019 Apr;23(8):3291-3301.doi: 10.26355/eurrev_201904_17691.Increased long noncoding RNA LINC00511 is correlated with poor prognosis and contributes to cell proliferation and metastasis by modulating miR-424 in hepatocellular carcinoma.长链非编码RNA LINC00511与肝细胞癌预后不良相关，并通过调节miR-424促进细胞增殖和转移

  104. ACS Sens.2019 Apr 26;4(4):977-983.doi: 10.1021/acssensors.9b00102. Epub 2019 Mar 21.Rapidly Visualizing the Membrane Affinity of Gene Vectors Using Polydiacetylene-Based Allochroic Vesicles.(IF:7.711).

  105. Gene.2019 Mar10;688:98-106. doi:10.1016/j.gene.2018.11.095.
    Embryonic morphology observation and HOXC8 gene expression in crest cushions of Chinese Crested duck

  106. J Cell Physiol . 2018 Jul;233(7):5405-5419. doi: 10.1002/jcp.26374. Epub 2018 Jan 23. Mechanical unloading reduces microtubule actin crosslinking factor 1 expression to inhibit β-catenin signaling and osteoblast proliferation

  107. Oncol Lett . 2018 Apr;15(4):4344-4350. doi: 10.3892/ol.2018.7831. Epub 2018 Jan 19. Synergistic effect of receptor-interacting protein 140 and simvastatin on the inhibition of proliferation and survival of hepatocellular carcinoma cells

  108. Toxicol Appl Pharmacol . 2017 May 1;322:75-88. doi: 10.1016/j.taap.2017.03.003. Epub 2017 Mar 8. Molecular mechanisms of 3,3’4,4′,5-pentachlorobiphenyl-induced epithelial-mesenchymal transition in human hepatocellular carcinoma cells

  109. Mol Med Rep . 2016 Feb;13(2):1725-31. doi: 10.3892/mmr.2015.4704. Epub 2015 Dec 22. Myricetin inhibits Kv1.5 channels in HEK293 cells.

  110. Nat Commun . 2015 Aug 24;6:8022. Rhomboid domain containing 1 promotes colorectal cancer growth through activation of the EGFR signalling pathway. (IF:11.329).

  111. Mol Cell Biol . 2015 Jul;35(14):2400-13. doi: 10.1128/MCB.00090-15. Epub 2015 May 4. SOCS3 Drives Proteasomal Degradation of TBK1 and Negatively Regulates Antiviral Innate Immunity

  112. Oncotarget . 2015 May 30;6(15):13049-59. Wedelolactone disrupts the interaction of EZH2-EED complex and inhibits PRC2-dependent cancer. (IF:6.359).

  113. Eur J Med Chem . 2014 Oct 6;85:119-26. doi: 10.1016/j.ejmech.2014.07.091. Epub 2014 Jul 25. Discovery of highly potent TNFα inhibitors using virtual screen.

  114. Biochem Biophys Res Commun . 2014 Apr 4;446(2):523-8. doi: 10.1016/j.bbrc.2014.02.137. Epub 2014 Mar 12. eEF1Bγ is a positive regulator of NF-кB signaling pathway.

  115. Int J Nanomedicine . 2013;8:3805-16. doi: 10.2147/IJN.S48950. Epub 2013 Oct 2. Efficacy and safety of dendrimer nanoparticles with coexpression of tumor necrosis factor-α and herpes simplex virus thymidine kinase in gene radiotherapy of the human uveal melanoma OCM-1 cell line. (IF:6.400).

  116. J Immunol . 2013 Sep 1;191(5):2614-23. doi: 10.4049/jimmunol.1203143. Epub 2013 Aug 5. Enhancer of zeste homolog 2 is a negative regulator of mitochondria-mediated innate immune responses.

  117. Appl Microbiol Biotechnol . 2013 Jul;97(14):6301-14. doi: 10.1007/s00253-013-4787-z. Epub 2013 Mar 6. Design, expression and characterization of a novel coexpression system of two antiarthritic molecules.

  118. Heart Vessels . 2013 May;28(3):385-96. doi: 10.1007/s00380-012-0275-0. Epub 2012 Sep 1. Real-time tracking of adipose tissue-derived stem cells with injectable scaffolds in the infarcted heart.

  119. PLoS One . 2012;7(11):e49567. doi:10.1371/journal.pone.0049567. Epub 2012 Nov 19. The ubiquitin ligase TRAF6 negatively regulates the JAK-STAT signaling pathway by binding to STAT3 and mediating its ubiquitination.

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反向转染

适用于高通量筛选、贴壁较慢的细胞，以及希望省去”提前一天铺板”步骤的实验。其原理是先在培养板内形成核酸-试剂复合物，再将悬浮的新鲜细胞加入，使复合物与细胞同步沉降接触。

⑴将0.5-0.8μg核酸用25μl无血清稀释液（OPTI-MEM、150mM NaCl溶液或5%葡萄糖）稀释，混匀。

⑵将0.5-0.8μl EntransterTM-H4000用25μl无血清稀释液稀释，室温静置5分钟。

⑶将试剂稀释液缓慢加入到核酸稀释液中（注意顺序，是将转染试剂加入到核酸中，而不是相反），充分混匀（加样器吹吸8-10次或短暂涡旋3-5秒，避免长时间剧烈振荡），室温静置15-20分钟（不超过20分钟）。转染复合物制备完成。

⑷将转染复合物50μl直接加入空的培养孔，轻摇使其均匀铺展于孔底。

⑸用胰酶消化处于对数生长期的细胞，重悬于含血清的完全培养基中，按1.2-2×10⁵ cells/孔（24-well）的密度加入到含复合物的孔中，终体积500μl。

⑹前后或左右轻摇培养板（不可画圈，否则细胞会向中心聚集），放入培养箱。

⑺4-6h后细胞贴壁，可酌情更换为新鲜完全培养基。后续检测时间同正向转染。

注意事项

  1. 反向转染对细胞接种密度更敏感：细胞当天接种、当天转染，接种密度需直接达到正向转染时应有的水平（并补偿胰酶消化后的贴壁损耗），24孔板1.2-2×10⁵/孔，各容器接种量见下方完整用量表。
  2. 不可使用含抗生素的培养基重悬细胞，转染4-6h后换液时再加回抗生素。
  3. 复合物在孔内可室温放置最多30分钟，超过则效率下降，建议复合物制备好后立即接种细胞。
  4. 不适用于神经元等极脆弱的原代细胞——胰酶消化后再接种会显著降低存活率。
  5. 反向转染完整用量表（核酸与试剂用量与正向相同，新增接种细胞数列，下表 HEK293 细胞为例）：

培养容器

核酸量

EntransterTM-H4000用量

稀释液（每路）

复合物总体积

接种细胞数

终培养体积

96-well

0.05-0.1μg

0.05-0.1μl

10μl

20μl

3-5×10⁴

100μl

48-well

0.2-0.3μg

0.2-0.4μl

15μl

30μl

6-10×10⁴

200μl

24-well

0.5-0.8μg

0.5-0.8μl

25μl

50μl

1.2-2×10⁵

500μl

12-well

1-1.5μg

1-1.5μl

50μl

100μl

2.5-4×10⁵

1ml

6-well/35-mm

2-2.5μg

2-3μl

100μl

200μl

1.0-1.5×10⁶

2ml

6cm dish

4-5μg

4-6μl

200μl

400μl

2-3×10⁶

4-5ml

10cm dish

8-10μg

8-12μl

500μl

1ml

6-8×10⁶

8-10ml

15cm dish

20-25μg

20-30μl

1-1.25ml

2-2.5ml

1.5-2×10⁷

20-25ml

注：反向转染中细胞处于刚贴壁起步阶段，对试剂毒性较正向敏感。如出现细胞批量脱落或贴壁不良，按以下顺序排查：①试剂量取表中下限（如24-well改0.5μl）；②细胞接种密度降10-20%；③若仍毒性，转染前6-12h做血清饥饿预适应。

  1. EntransterTM系列转染试剂的成分和原理，化学结构，粒径，电位等数据

  2. siRNA转染与DNA转染有什么不一样?

  3. 为什么EntransterTM转染试剂有血清转染不仅没有毒性，反而有助于提高转染效率?

相关问题

siRNA和质粒共转染

本系列试剂分工不同：EntransterTM-R4000适用于siRNA等小片段RNA，EntransterTM-H4000适用于质粒DNA与长mRNA；siRNA与质粒共转染时建议分步进行——先用R4000转染siRNA，24小时后细胞状态良好（无需换液）再用H4000转染质粒DNA。应用场景与策略：

①基因敲低+不相关基因过表达：按上述分步操作即可。

②Rescue实验（敲低内源基因+过表达突变体）：质粒序列必须含silent同义突变避开siRNA识别区（通常在siRNA靶序列内的密码子第3碱基做3-5个沉默替换），否则plasmid同样会被敲低。

③不可做的组合：敲低内源基因+同时过表达无siRNA抗性的同基因plasmid——结果会被同步敲低，无法解读。

下表给出各类检测的推荐时间点（均自siRNA转染后计时）：

检测类型

推荐时间点（siRNA转染后）

说明

qPCR (mRNA)

12h / 24h / 48h / 72h

mRNA敲低6-12h已达80%以上，12h常达峰

Western (蛋白)

24h / 48h / 72h / 96h

短半衰期蛋白 24-48h 已见下降

报告基因 (荧光/荧光素酶)

24h / 48h / 72h

24h 初见，48h 高峰，72h 观察持续

功能实验：增殖

48-72h

蛋白耗竭后增殖差异渐显，48h起可读出

功能实验：迁移/侵袭

48-96h

划痕/Transwell周期较长，48h后读出更稳定

细胞毒性/活力

24h / 48h / 72h

捕捉早期与延迟毒性

注1：siRNA敲低效应通常可持续5-7天（化学修饰siRNA可达10天）。如检测窗口超出96h，建议补做120h、168h时间点。

注2：首次研究某基因时，建议做0/12/24/48/72/96h完整时间序列以确定该靶基因的knockdown高峰与维持窗口，后续实验只取最优时间点。

常见问题与解决方案

问题

可能原因

解决方案

完全无表达（任意细胞均零信号）

阳性对照不足或质粒构建缺陷

用已知阳性质粒（如pEGFP-N1）排查；检查启动子在该细胞系活性（CMV在原代细胞效率低）；mRNA则检查IVT产物完整性

完全无表达

稀释液错误或被污染

必须用OPTI-MEM、150mM NaCl或5%葡萄糖；严禁使用PBS（磷酸根使试剂沉淀）、含血清或含双抗的培养基

转染效率<10%

细胞代数过高或处于衰老期

改用20代以内的细胞，确保对数生长期（无饥饿/过度融合/老化）

核酸纯度不足

参考二节核酸纯度要求：OD260/280=1.8-2.0、OD260/230>2.0、内毒素<0.1EU/μg、浓度>500ng/μl，无乙醇和盐残留

转染效率20-50%（可优化）

试剂:核酸比未达最优

做0.5:1/1:1/1.5:1 (μl:μg)三点梯度筛选（参考表1按容器对应比例）；首次使用某细胞系建议做3×3 DoE：试剂:核酸 (μl:μg) 0.5:1/1:1/1.5:1 × 细胞密度（贴壁50/65/80%汇合）；系统优化另见三节

转染后细胞批量脱落漂浮

试剂过量或双抗协同毒性

试剂量降至1:1 (μl:μg)以下（即每μg核酸用1μl试剂以下）；转染当下及前4-6h使用无双抗培养基，4-6h换液时再加回双抗

转染后24-48h细胞缓慢凋亡

核酸含内毒素

改用Endo-Free级试剂盒提取；目标值<0.1EU/μg；用于难转染或敏感细胞时<0.01EU/μg

排除以上仍有毒性

质粒表达产物本身具毒性

用pEGFP-N1等已知低毒质粒做阳性对照对比，区分是试剂问题还是构建问题

复合物制备后出现絮状或沉淀

稀释液污染（PBS、血清、蛋白、双抗）

改用新鲜OPTI-MEM、150mM NaCl或5%葡萄糖重做；核酸与试剂稀释液严禁含血清/双抗

复合物呈乳白色浑浊

孵育时间>20min或核酸浓度过高

缩短至15min；增大稀释体积降低核酸浓度（参考表1的稀释液体积）

mRNA转染零信号

IVT mRNA缺cap或poly(A)结构缺陷

检查5’cap (m⁷G/ARCA/CleanCap)与3’poly(A)长度≥120nt；用denaturing胶或Bioanalyzer验证完整性；DNase处理去除模板残留

mRNA表达短暂（<6h即消失）

RNase污染或未修饰mRNA易降解

全程RNase-free操作（DEPC水、新tip、专用稀释液）；改用N1-methyl-pseudouridine (m1Ψ) 修饰mRNA延长半衰期

mRNA转染细胞毒性大

dsRNA杂质或未修饰mRNA触发先天免疫应答

用m1Ψ修饰mRNA；HPLC或纤维素柱纯化去除dsRNA；首次使用前用低浓度试做

长mRNA (>3kb) 转染效率显著低

复合物聚集加快、内吞效率下降

mRNA复合物孵育缩短至5-10min；试剂量上调至DNA推荐量的1.5倍；可尝试离心法增加接触

反向转染细胞结团或贴壁不均

加细胞后画圈摇板

改为前后或左右轻柔摇动培养板；严禁画圈，否则细胞向孔中心聚集

反向转染效率显著低于正向

复合物在空孔内放置过久

复合物制备完成后30min内必须接种细胞；超过30min效率显著下降，需重做

siRNA与DNA共转染表达水平差

同步转染时两者相互干扰

改用分步法：先用EntransterTM-R4000转siRNA→24h后用EntransterTM-H4000转DNA，参考五节

采取"血清饥饿"方法提高转染效率

对常见永生化贴壁细胞，如HeLa、HEK293/HEK293T、NIH3T3、Cos-7、A549、MCF-7以及更依赖血清的癌细胞，如PC12、SH-SY5Y、HepG2、MDCK等，还可以采取”血清饥饿”方法提高转染效率。转染前6-12h，采用无血清或低血清（1%）培养基培养细胞，转染前更换为含正常血清含量如10%的培养基，然后转染。注意此方法不适用于原代细胞、已处于弱增殖或分化状态的细胞。

采用“离心法”提高转染效率。

对悬浮细胞、难转染的原代细胞(原代 T 细胞、B 细胞、骨髓源性巨噬细胞 (BMDM)、神经元等)，病毒包装、CRISPR 基因编辑等还可以采用“离心法”以增大转染复合物与细胞的接触，提高转染效率。转染后用封口膜严密包裹培养板边缘，配平后室温下以500-1000g使用带水平转子（Swing-bucket rotor）的离心机离心30-60min，离心完成后撕去封口膜，酒精擦拭板外壁。将培养板放入培养箱中继续孵育。

运输与储存

常温运输，于4℃长期存储，有效期12个月。

安全性

本品使用安全，未发现任何生物、化学毒性。如不慎沾染，用清水冲洗即可。

质量保证

北京英格恩生物科技有限公司本产品实行严格质量检验，并进行转染验证，以确保产品质量。请用户使用前务必认真阅读操作手册。如用户仍然有问题，请立即联系本公司人员，英格恩公司将负责提供全程技术服务。

使用限制

本品仅限科研用途。

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