Lonza Nucleofector 2b單孔細(xì)胞核轉(zhuǎn)染系統(tǒng)
——原代細(xì)胞、難轉(zhuǎn)染細(xì)胞系高效轉(zhuǎn)染解決方案

 
北京澤平代理的德國(guó)進(jìn)口Lonza Nucleofector 2b單孔細(xì)胞核轉(zhuǎn)染系統(tǒng)（原Amaxa Nucleofector II/2b Device，又稱Lonza 2b細(xì)胞核轉(zhuǎn)儀、Lonza 2b細(xì)胞電轉(zhuǎn)儀）， 作為全球知名的高效基因電轉(zhuǎn)儀，其利用傳統(tǒng)電穿孔原理，結(jié)合細(xì)胞特異性電轉(zhuǎn)染液，可以將包括DNA、RNA、質(zhì)粒、多肽、蛋白質(zhì)、核糖核蛋白復(fù)合體RNP、小分子化合物等轉(zhuǎn)入細(xì)胞質(zhì)中，并穿過核膜直接進(jìn)入細(xì)胞核中，實(shí)現(xiàn)高效轉(zhuǎn)染，稱為Nucleofector核轉(zhuǎn)染技術(shù)(Nucleofection)。其中質(zhì)粒DNA轉(zhuǎn)染效率最高達(dá)到90%、寡核苷酸如siRNA轉(zhuǎn)染效率最高達(dá)到99%。


 

Lonza 2b電轉(zhuǎn)儀自2001年上市以來(lái)，廣泛用于包括干細(xì)胞、神經(jīng)細(xì)胞、T淋巴細(xì)胞在內(nèi)的動(dòng)物原代細(xì)胞和難轉(zhuǎn)染的細(xì)胞系，以及細(xì)菌、外泌體轉(zhuǎn)染中，助力CAR-T、CAR-NK、CRISPR/Cas9基因編輯、IPS重編程、RNA干擾、外泌體遞送等多種前沿研究，全球發(fā)表文章超過14,000篇。



 

2b電轉(zhuǎn)儀不依賴于有絲分|裂，尤其適合不分化細(xì)胞，如靜止的T淋巴細(xì)胞、神經(jīng)細(xì)胞等，最快2小時(shí)可觀察到蛋白表達(dá)。相比脂質(zhì)體轉(zhuǎn)染、病毒轉(zhuǎn)導(dǎo)、傳統(tǒng)電穿孔儀，Lonza 2b電轉(zhuǎn)儀操作簡(jiǎn)單，重復(fù)性高，在原代細(xì)胞和細(xì)胞系中實(shí)現(xiàn)更高轉(zhuǎn)染效率、更高細(xì)胞活率、更快基因表達(dá)，加快推進(jìn)實(shí)驗(yàn)進(jìn)程。


Fig. Primary NHDF-neo cells were transfected with labeled plasmid DNA encoding GFP, fixed after 2h in 3.5%PFA and analyzed by confocal microscopy.
 

 

Lonza Nucleofector核轉(zhuǎn)染技術(shù)依托電轉(zhuǎn)儀設(shè)備、電轉(zhuǎn)染試劑盒、全球共享數(shù)據(jù)庫(kù)三方面技術(shù)和資源優(yōu)勢(shì)，實(shí)現(xiàn)高效、靈活、方便的細(xì)胞轉(zhuǎn)染體驗(yàn)。



①2b電轉(zhuǎn)儀——內(nèi)置針對(duì)不同細(xì)胞類型優(yōu)化的電脈沖程序，無(wú)需人工設(shè)置電壓電流等電擊參數(shù)，無(wú)需摸索轉(zhuǎn)染條件，選定程序一鍵操作，實(shí)驗(yàn)過程簡(jiǎn)單方便。

②電轉(zhuǎn)染試劑盒——Lonza 2b電轉(zhuǎn)染試劑盒由電極杯（電轉(zhuǎn)杯）、電轉(zhuǎn)染緩沖液、補(bǔ)充劑、pmaxGFP陽(yáng)性對(duì)照質(zhì)粒、巴氏吸管組成。其電轉(zhuǎn)緩沖液配方根據(jù)原代細(xì)胞、細(xì)胞系類型單獨(dú)優(yōu)化，每種試劑盒用于不同細(xì)胞類型，以提高轉(zhuǎn)染效率、轉(zhuǎn)染后細(xì)胞活率，支持多種底物共轉(zhuǎn)染，實(shí)驗(yàn)結(jié)果具備高重復(fù)性。

③全球共享數(shù)據(jù)庫(kù)——線上實(shí)時(shí)數(shù)據(jù)庫(kù)（https://knowledge.lonza.com），可查詢超過759種細(xì)胞的轉(zhuǎn)染全流程數(shù)據(jù)，包括轉(zhuǎn)染前細(xì)胞來(lái)源、傳代、培養(yǎng)條件、轉(zhuǎn)染程序選擇和操作技巧、轉(zhuǎn)染后培養(yǎng)條件、歷史轉(zhuǎn)染結(jié)果等，資源不斷更新，豐富便捷的數(shù)據(jù)參考，提高實(shí)驗(yàn)效率，幫助科研人員專注于研究本身。

 

 

Lonza 2b細(xì)胞電轉(zhuǎn)儀轉(zhuǎn)染操作流程



 

Lonza 2b電轉(zhuǎn)儀參數(shù)
 

 
引用文獻(xiàn)

1. Korbinian N. Kropp, et al.(2023) Targeting the melanoma-associated antigen CSPG4 with HLA-C*07:01-restricted T-cell receptors. https://doi.org/10.3389/fimmu.2023.1245559
2. Stephan Riesenberg, et al.(2023) Efficient high-precision homology-directed repair-dependent genome editing by HDRobust. https://doi.org/10.1038/s41592-023-01949-1
3. Kui Zhao, et al.(2023) Generation of an NSD2-deficient human embryonic stem cell line using CRISPR/Cas9 technology. https://doi.org/10.1016/j.scr.2023.103255
4. Meiling Jiang, et al.(2023) Generation of a homozygous RANGRF knockout hiPSC line by CRISPR/Cas9 system. https://doi.org/10.1016/j.scr.2023.103136
5. Xingjie Ren, et al.(2023) Efficient bi-allelic tagging in human induced pluripotent stem cells using CRISPR. https://doi.org/10.1016/j.xpro.2023.102084
6. Ittetsu Nakajima, et al.(2023) In Vivo Delivery of Therapeutic Molecules by Transplantation of Genome-Edited Induced Pluripotent Stem Cells. https://doi.org/10.1177/09636897231173734
7. Shidong Qiu, et al.(2023) Generation of the induced pluripotent stem cell line SFMUi001-A from a patient with usher syndrome type 2 caused by biallelic variants in the USH2A gene. https://doi.org/10.1016/j.scr.2023.103101
8. Chiami Moyama, et al.(2023) Myb Repression Mediates Stat5b-knockdown-induced Apoptosis and Inhibits Proliferation of Glioblastoma Stem Cells. https://doi.org/10.21873/cgp.20374
9. Saito, M.K., et al.(2023) A disease-specific iPS cell resource for studying rare and intractable diseases.  https://doi.org/10.1186/s41232-023-00294-2
10. Joseph T. Smith, et al.(2023) Developmental dynamics of mitochondrial mRNA abundance and editing reveal roles for temperature and the differentiation-repressive kinase RDK1 in cytochrome oxidase subunit II mRNA editing. https://doi.org/10.1128/mbio.01854-23

 


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——Lonza中國(guó)一級(jí)代理商，北京澤平

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