TY - JOUR
T1 - Systematic selection of housekeeping genes for gene expression normalization in chicken embryo fibroblasts infected with Newcastle disease virus
AU - Yin, Renfu
AU - Liu, Xinxin
AU - Liu, Chang
AU - Ding, Zhuang
AU - Zhang, Xiaodong
AU - Tian, Furong
AU - Liu, Weihong
AU - Yu, Jinghai
AU - Li, Lin
AU - Hrabé de Angelis, Martin
AU - Stoeger, Tobias
PY - 2011/10/7
Y1 - 2011/10/7
N2 - Gene expression analysis is frequently used to analyze the response to viral infection, and 18S RNA, SHDA and GAPDH represent popular house keeping genes (HKGs) often used to normalize gene expression. Here we describe the first systematic selection and evaluation of suitable HKGs for gene expression analysis in chicken embryo fibroblasts (CEF) infected with NDV adapted to the guidelines from Gorzelniak and Ferguson. Our results indicate that ACTB, HPRT1 and HMBS were valuable and stable HKGs, while 18S RNA, GAPDH and SHDA are considerably regulated during the course of infection and thus precluded for normalization. Normalizing the infection dependent gene IFN-a and the infection independent gene B2M to inappropriate HKGs consequently misleads to significant errors in estimating their regulations. Our study emphasizes that even the most popular HKGs like 18S RNA and GAPDH can lead to divergent and inaccurate data interpretation of significant magnitude if not carefully analyzed for stability before.
AB - Gene expression analysis is frequently used to analyze the response to viral infection, and 18S RNA, SHDA and GAPDH represent popular house keeping genes (HKGs) often used to normalize gene expression. Here we describe the first systematic selection and evaluation of suitable HKGs for gene expression analysis in chicken embryo fibroblasts (CEF) infected with NDV adapted to the guidelines from Gorzelniak and Ferguson. Our results indicate that ACTB, HPRT1 and HMBS were valuable and stable HKGs, while 18S RNA, GAPDH and SHDA are considerably regulated during the course of infection and thus precluded for normalization. Normalizing the infection dependent gene IFN-a and the infection independent gene B2M to inappropriate HKGs consequently misleads to significant errors in estimating their regulations. Our study emphasizes that even the most popular HKGs like 18S RNA and GAPDH can lead to divergent and inaccurate data interpretation of significant magnitude if not carefully analyzed for stability before.
KW - Chicken embryo fibroblasts
KW - Housekeeping gene
KW - Newcastle disease virus
KW - QRT-PCR
UR - http://www.scopus.com/inward/record.url?scp=80053583557&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2011.08.131
DO - 10.1016/j.bbrc.2011.08.131
M3 - Article
C2 - 21925148
AN - SCOPUS:80053583557
SN - 0006-291X
VL - 413
SP - 537
EP - 540
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 4
ER -