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浙江大學高效率轉染質粒DNA、siRNA到免疫T(Vδ1 T)細胞,發表文章到nature-sigtrans

發布:2021-04-08 21:25:23 瀏覽:526次

一、浙江大學附屬第二醫院乳腺外科;浙江大學附屬第二醫院浙江省療法腫瘤微環境與免疫重點實驗室;浙江省人民醫院,浙江省人民醫院,杭州醫學院附屬人民醫院,浙江省個體化醫學腫瘤分子診斷與診斷重點實驗室;紹興大學附屬醫院甲狀腺乳腺外科聯合發表標題為Breast cancer-derived exosomes transmit lncRNA SNHG16 to induce CD73+γδ1 Treg cells(乳腺癌衍生的外泌體傳遞lncRNA SNHG16誘導CD73 +γδ1 Treg細胞)的文章到nature.com/sigtrans,文章已被接受;

二、本研究使用中使用Invigentech(英克)公司INVI DNA RNA 轉染試劑轉染質粒DNA、siRNA到Vδ1 T 免疫T細胞里面;

三、本文中轉染質粒DNA、siRNA,轉染細胞數量信息如下:

A. 將全長2435 bp的序列克隆到pCR3.1載體中構建SNHG16過表達載體;

B. siRNA序列:SNHG16-homo-349,5′GCCUCUGCUGCUAAUUGUUTT-3'; SNHG16-homo-867, 5′-CCAAGGAGGGACUGUUUAATT-3′和SNHG16-homo-2004,5′-CCCAGUGUUGACUCACCAATT-3;

C. 轉染細胞用量:6孔板里面1 × 106 cells/well;

四、發表文章部分內容如下:

Breast cancer-derived exosomes transmit lncRNA SNHG16 to induce CD73+γδ1 Treg cells

Plasmid construction and siRNA silencing A full-length 2435 bp sequence was cloned into the pCR3.1 vector to

construct the SNHG16 overexpression vector. The sequences of SNHG16-specifific siRNAs were as follows: SNHG16-homo-349, 5′GCCUCUGCUGCUAAUUGUUTT-3′; SNHG16-homo-867, 5′-CCAAGGAGGGACUGUUUAATT-3′ and SNHG16-homo-2004, 5′-CCCAGUGUUGACUCACCAATT-3′. The miR-16–5p mimics, inhibitor and negative

controls were purchased from GenePharma (SupplementaryTable S3).

To manipulate the expression of miR-16–5p in Vδ1 T cells, Vδ1T cells were sorted from peripheral blood and seeded into six-wellplates at 1 × 106 cells/well with 1 ml of medium supplementedwith 10% FBS, 10 μg/ml CD3, 10 μg/ml CD28 and 40 U/mL IL-2.Then, transfection reagent (INVI DNA RNA Transfection Reagent,Invigentech, USA) and miR-16–5p mimics/NC/inhibitor/inhibitor NC (20 μm) were incubated with the cells for 48 h, after which the cells were collected for further experiments.

but not Vδ2+ T cells were revealed to comprise the majority of TILs in all three BC molecular subtypes when gated on CD3 (p < 0.01, Fig. 1b, c). In addition, immunoflfluorescence of frozen sections further confifirmed the dominant distribution of Vδ1+T cells in BC (Fig. 1d). 

To further identify the function of breast cancer-infifiltrating γδ1T cells, BC cell lines (MCF-7, T-47D, MDA-MB-231 and MDA-MB-468) were co-cultured with freshly isolated Vδ1+ T cells frombreast tumour tissue (E:T ratios: 1:1, 5:1 and 10:1). Upon plotting the cell growth curve, the data show that γδ1 T cells did not exert

invigentech(英克)INVI DNA RNA轉染試劑信息如下

產品名稱貨號規格報價
INVI DNA RNA 轉染試劑IV12160250.25 ml

詢價

INVI DNA RNA 轉染試劑IV12160500.50 ml

詢價

INVI DNA RNA 轉染試劑IV12160750.75 ml詢價
INVI DNA RNA 轉染試劑IV12161001.00 ml詢價
INVI DNA RNA 轉染試劑IV12161501.50 ml詢價
INVI DNA RNA 轉染試劑IV12163003.00 ml詢價

www.ivigt.com


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