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技术文章
浙江大学使用Invigentech（英克）公司INVI DNA RNA转染试剂高效率转染质粒DNA siRNA到免疫T（Vδ1 T）细胞
点击次数：5522 发布时间：2020/5/7 16:41:09
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在2020-2-19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.
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 | IV1216025 | 0.25 ml | 询价 |
| INVI DNA RNA 转染试剂 | IV1216050 | 0.50 ml | 询价 |
| INVI DNA RNA 转染试剂 | IV1216075 | 0.75 ml | 询价 |
| INVI DNA RNA 转染试剂 | IV1216100 | 1.00 ml | 询价 |
| INVI DNA RNA 转染试剂 | IV1216150 | 1.50 ml | 询价 |
| INVI DNA RNA 转染试剂 | IV1216300 | 3.00 ml | 询价 |
原创作者：上海徕同生物科技有限公司
