| Home  | About ScienceAsia  | Publication charge  | Advertise with us  | Subscription for printed version  | Contact us  
Editorial Board
Journal Policy
Instructions for Authors
Online submission
Author Login
Reviewer Login
Volume 49 Number 1
Volume 48 Number 6
Volume 48 Number 5
Volume 48 Number 4
Volume 48 Number 3
Volume 48 Number 2
Earlier issues
Volume 40 Number 4 Volume 40 Number 5 Volume 40 Number 6

previous article next article

Research articles

ScienceAsia 40 (2014): 340-347 |doi: 10.2306/scienceasia1513-1874.2014.40.340

RNAi down-regulation of Nck1 adaptor protein in Jurkat T cells

Pussadee˙Paensuwana, Jatuporn˙Ngoenkama, Donruedee˙Sanguansermsria, Boonruang˙Khamsria, Ichaya˙Yiemwattanab, Apirath˙Wangteeraprasertc, Sutatip˙Pongcharoenc,d,*

ABSTRACT:     RNA interference (RNAi) is a potent gene delivery system for studying the regulation of gene expression in a wide variety of eukaryotic cells. In the present study, different RNAi approaches, namely, synthetic small interfering RNA (siRNA) and plasmid- and lentivirus-based short hairpin RNA (shRNA) were investigated to assess the down-regulation of Nck1 protein efficiency in the Jurkat T cell line. Jurkat T cells treated with these three different systems substantially and specifically reduced the expression of the Nck1 protein but not that of the Nck2 protein. Although the three systems showed a similar Nck1 knockdown efficiency, they led to different T cell activation outcomes. After stimulation, CD69 expression and IL-2 production were impaired in Nck1-siRNA and plasmid-based Nck1-shRNA transfected Jurkat cells. However, these T cell activation outcomes were increased in lentiviral vector based Nck1-shRNA transfected cells. These data suggest that the outcomes from transfection with the shRNA based lentiviral vector differ from those of siRNA and shRNA-based plasmids although they provide the same gene silencing efficiency. The verification of suitable RNAi strategies to silence target genes is therefore necessary before using it in experiments.

Download PDF

12 Downloads 1189 Views

a Department˙of˙Microbiology˙and˙Parasitology, Faculty˙of˙Medical˙Science, Naresuan˙University, Pitsanulok˙65000˙Thailand
b Department˙of˙Preventive˙Dentistry, Faculty˙of˙Dentistry, Naresuan˙University, Pitsanulok˙65000˙Thailand
c Department˙of˙Medicine, Faculty˙of˙Medicine, Naresuan˙University, Phitsanulok˙65000˙Thailand
d Centre˙of˙Excellence˙in˙Medical˙Biotechnology, Faculty˙of˙Medical˙Science, Naresuan˙University, Pitsanulok˙65000˙Thailand

* Corresponding author, E-mail: sutatipp@nu.ac.th

Received 25 Apr 2014, Accepted 29 Sep 2014