GapmeR-Mediated Gene Silencing in Motile T-Cells
Gene silencing is an important method to study gene functions in health and diseases. While there are various techniques that are applied to knockdown specific gene(s) of interest, they have certain limitations in application to T-lymphocytes. T-cells are “hard-to-transfect” cells and are recalcitrant to transfection reagents. Here, we describe the use of novel cell-permeating antisense molecules, called “GapmeR”, to knockdown specific gene(s) in human primary T-cells.
Key wordsGapmeR RNAi Gene knockdown T-Lymphocytes
This work was supported in part by grants from Lee Kong Chian School of Medicine, Nanyang Technological University Singapore Start-Up Grant to N.K.V. and the Singapore Ministry of Education under its Singapore Ministry of Education (MOE) Academic Research Fund (AcRF) Tier 2 Grant (MOE2017-T2-2-004). A.K. acknowledges Ph.D. fellowship provided by Lee Kong Chian School of Medicine, Nanyang Technological University Singapore.
- 7.Stein CA, Hansen JB, Lai J, Wu S, Voskresenskiy A, Høg A, Worm J, Hedtjärn M, Souleimanian N, Miller P, Soifer HS, Castanotto D, Benimetskaya L, Ørum H, Koch T (2010) Efficient gene silencing by delivery of locked nucleic acid antisense oligonucleotides unassisted by transfection reagents. Nucleic Acids Res 38:e3CrossRefGoogle Scholar
- 8.Seth PP, Siwkowski A, Allerson CR, Vasquez G, Lee S, Prakash TP, Wancewicz EV, Witchell D, Swayze EE (2009) Short antisense oligonucleotides with novel 2′-4′ conformationaly restricted nucleoside analogues show improved potency without increased toxicity in animals. J Med Chem 52:10–13CrossRefGoogle Scholar
- 10.Ong ST, Chalasani MLS, Fazil MHUT, Praseetha P, Kizhakeyil A, Wright GD, Kelleher D, Verma NK (2018) Centrosome- and Golgi-localized protein kinase N-associated protein serves as a docking platform for protein kinase A signaling and microtubule nucleation in migrating T-cells. Front Immunol 9:397CrossRefGoogle Scholar
- 11.Verma NK, Dempsey E, Conroy J, Olwell P, Mcelligott AM, Davies AM, Kelleher D, Butini S, Campiani G, Williams DC, Zisterer DM, Lawler M, Volkov Y (2008) A new microtubule-targeting compound PBOX-15 inhibits T-cell migration via post-translational modifications of tubulin. J Mol Med 86:457–469CrossRefGoogle Scholar