A fast, simple method for screening radiation susceptibility genes by RNA interference.
Radiotherapy can cause unacceptable levels of damage to normal tissues in some cancer patients. To understand the molecular mechanisms underlying radiation-induced physiological responses, and to be able to predict the radiation susceptibility of normal tissues in individual patients, it is important to identify a comprehensive set of genes responsible for radiation susceptibility. We have developed a simple and rapid 96-well screening protocol using cell proliferation assays and RNA interference to identify genes associated with radiation susceptibility. We evaluated the performance of alamarBlue-, BrdU-, and sulforhodamine B-based cell proliferation assays using the 96-well format. Each proliferation assay detected the known radiation susceptibility gene, PRKDC. In a trial screen using 28 shRNA vectors, another known gene, CDKN1A, and one new radiation susceptibility gene, ATP5G3, were identified. Our results indicate that this method may be useful for large-scale screens designed to identify novel radiation susceptibility genes.
Tsuji
Atsushi B
AB
RadGenomics Project, Frontier Research Center, National Institute of Radiological Sciences, Chiba, Japan.
Sudo
Hitomi
H
Sugyo
Aya
A
Otsuki
Marika
M
Miyagishi
Makoto
M
Taira
Kazunari
K
Imai
Takashi
T
Harada
Yoshi-Nobu
YN
D002452:Cell Count / Q000379:methods*; D002470:Cell Survival / Q000528:radiation effects*; D004198:Disease Susceptibility / Q000175:diagnosis; D004305:Dose-Response Relationship, Drug; D020869:Gene Expression Profiling / Q000379:methods*; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D034622:RNA Interference; D011829:Radiation Dosage; D011832:Radiation Injuries / Q000175:diagnosis / Q000209:etiology / Q000235:genetics*; D011836:Radiation Tolerance / Q000235:genetics*; D011839:Radiation, Ionizing; D018570:Risk Assessment / Q000379:methods*; D012307:Risk Factors