浙江大学高效率转染质粒DNA、siRNA到免疫T(Vδ1 T)细胞,发表文章到nature-sigtrans

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浙江大学高效率转染质粒DNA、siRNA到免疫T(Vδ1 T)细胞,发表文章到nature-sigtrans
浙江大学高效率转染质粒DNA、siRNA到免疫T(Vδ1 T)细胞,发表文章到nature-sigtransimage.png

一、浙江大学附属第二医院乳腺外科;浙江大学附属第二医院浙江省疗法肿瘤微环境与免疫重点实验室;浙江省人民医院,浙江省人民医院,杭州医学院附属人民医院,浙江省个体化医学肿瘤分子诊断与诊断重点实验室;绍兴大学附属医院甲状腺乳腺外科联合发表标题为Breast cancer-derived exosomes transmit lncRNA SNHG16 to induce CD73+γδ1 Treg cells(乳腺癌衍生的外泌体传递lncRNA SNHG16诱导CD73 +γδ1 Treg细胞)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 转染试剂转染质粒DNA、siRNA到Vδ1 T 免疫T细胞里面;

三、本文中转染质粒DNA、siRNA,转染细胞数量信息如下:

A. 将全长2435 bp的序列克隆到pCR3.1载体中构建SNHG16过表达载体;

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

C. 转染细胞用量:6孔板里面1 × 106 cells/well;

四、发表文章部分内容如下:

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.

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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转染试剂信息如下

产品名称货号规格报价
INVI DNA RNA 转染试剂IV12160250.25 ml

询价

INVI DNA RNA 转染试剂IV12160500.50 ml

询价

INVI DNA RNA 转染试剂IV12160750.75 ml询价
INVI DNA RNA 转染试剂IV12161001.00 ml询价
INVI DNA RNA 转染试剂IV12161501.50 ml询价
INVI DNA RNA 转染试剂IV12163003.00 ml询价

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