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浙江大学使用Invigentech（英克）INVI DNA RNA 转染试剂高效率转染质粒DNA、siRNA到免疫T（Vδ1 T）细胞，发表文章到nature-sigtrans

阅读次数：2523 发布时间：2020/5/7 16:39:27

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