浙江大学科研者告知您 如何高效率转染免疫T（Vδ1 T）细胞

高效率转染免疫T细胞是大家关心的问题；

一、2020-2-19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；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杂志，文章已被接受；

二、本研究使用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、DNA片段大小等信息如下：

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.

invigentech（英克）INVI DNA RNA转染试剂信息如下

www.ivigt.com