Use of GapmeRs for gene expression knockdowns in human primary resting CD4+T cells

被引：2

作者：

Abewe, Hosiana ^{[1
,2
,3
,4
]}

Deshmukh, Savitha ^{[1
,2
]}

Mukim, Amey ^{[1
,2
]}

Beliakova-Bethell, Nadejda ^{[1
,2
,3
]}

机构：

[1] San Diego VA Med Ctr, San Diego, CA 92161 USA

[2] Vet Med Res Fdn, San Diego, CA 92161 USA

[3] Univ Calif San Diego, Stein Clin Res Bldg,Rm 304,9500 Gilman Dr,0679, La Jolla, CA 92093 USA

[4] Univ Utah, Sch Med, Salt Lake City, UT 84112 USA

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2020年 / 476卷

基金：

美国国家卫生研究院;

关键词：

Gene expression; Knockdown; Antisense oligonucleotides; GapmeRs; Primary CD4+T cells; Resting CD4+T cells; TRANSCRIPTOME;

D O I：

10.1016/j.jim.2019.112674

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页数：6

共 14 条

[1] Histone deacetylase inhibitors induce complex host responses that contribute to differential potencies of these compounds in HIV reactivation [J].