Human primary resting CD4+ T cells are difficult to transfect while preserving viability. The present study evaluated gymnotic delivery and RNase H1-dependent gene expression knockdown mediated by antisense oligonucleotides, called GapmeRs. Exposure of primary resting CD4+ T cells to GapmeRs did not cause cell activation or affect cell viability. Gene expression knockdowns were stable at least up to 48 h after removal of GapmeRs from culture. Exposure to GapmeRs resulted in comparable levels of degradation along the entire transcript, which could be important when studying function of regulatory long non-coding RNAs. Efficiency of transcript degradation was not solely dependent on the dose of GapmeR, RNA target and its localization. When using GapmeRs, some optimization is required, and all targets have to be individually tested; however, using GapmeRs is advantageous in experiments where preservation of the resting state of the human primary CD4+ T cells and targeting nuclear RNAs are desired. In certain cases, combining GapmeR with siRNA for the same target may improve knockdown efficiency.

中文翻译：

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使用 GapmeR 敲低人类原代静息 CD4+ T 细胞的基因表达。


人类原代静息 CD4+ T 细胞很难在保持活力的同时进行转染。本研究评估了反义寡核苷酸（称为 GapmeR）介导的体操递送和 RNase H1 依赖性基因表达敲低。将原代静息 CD4+ T 细胞暴露于 GapmeR 不会导致细胞激活或影响细胞活力。从培养物中去除 GapmeR 后，基因表达敲低至少可以稳定长达 48 小时。暴露于 GapmeR 会导致整个转录本出现类似水平的降解，这在研究调节性长非编码 RNA 的功能时可能很重要。转录物降解效率不仅仅取决于 GapmeR、RNA 靶标及其定位的剂量。使用GapmeRs时，需要进行一些优化，并且所有目标都必须单独测试；然而，在需要保存人类原代 CD4+ T 细胞静息状态并靶向核 RNA 的实验中，使用 GapmeR 是有利的。在某些情况下，针对同一靶标将 GapmeR 与 siRNA 组合可能会提高敲低效率。