Inhibition of herpes simplex virus 1 gene expression and replication by RNase P-associated external guide sequences.
An external guide sequence (EGS) is a RNA sequence which can interact with a target mRNA to form a tertiary structure like a pre-tRNA and recruit intracellular ribonuclease P (RNase P), a tRNA processing enzyme, to degrade target mRNA. Previously, an in vitro selection procedure has been used by us to engineer new EGSs that are more robust in inducing human RNase P to cleave their targeted mRNAs. In this study, we constructed EGSs from a variant to target the mRNA encoding herpes simplex virus 1 (HSV-1) major transcription regulator ICP4, which is essential for the expression of viral early and late genes and viral growth. The EGS variant induced human RNase P cleavage of ICP4 mRNA sequence 60 times better than the EGS generated from a natural pre-tRNA. A decrease of about 97% and 75% in the level of ICP4 gene expression and an inhibition of about 7,000- and 500-fold in viral growth were observed in HSV infected cells expressing the variant and the pre-tRNA-derived EGS, respectively. This study shows that engineered EGSs can inhibit HSV-1 gene expression and viral growth. Furthermore, these results demonstrate the potential for engineered EGS RNAs to be developed and used as anti-HSV therapeutics.
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.
Jin
J
Liu
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.
Luyao
L
Shao
School of Public Health, University of California, Berkeley, CA 94720, USA.
Phong
P
Trang
Taizhou Institute of Virology, Taizhou, Jiangsu 225300, China.|Jiangsu Affynigen Biotechnologies, Inc., Taizhou, Jiangsu 225300, China.
Zhu
Z
Yang
School of Public Health, University of California, Berkeley, CA 94720, USA.
Michael
M
Reeves
College of Life Sciences, Jinan University, Guangzhou, Guangdong 510632, China.
Xu
X
Sun
School of Public Health, University of California, Berkeley, CA 94720, USA.
Gia-Phong
GP
Vu
Taizhou Institute of Virology, Taizhou, Jiangsu 225300, China.|Jiangsu Affynigen Biotechnologies, Inc., Taizhou, Jiangsu 225300, China.|College of Life Sciences, Jinan University, Guangzhou, Guangdong 510632, China.
Yu
Y
Wang
College of Life Sciences, Jinan University, Guangzhou, Guangdong 510632, China.
Hongjian
H
Li
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.
Congyi
C
Zheng
School of Public Health, University of California, Berkeley, CA 94720, USA.
Sangwei
S
Lu
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.|School of Public Health, University of California, Berkeley, CA 94720, USA.|College of Life Sciences, Jinan University, Guangzhou, Guangdong 510632, China.
Fenyong
F
Liu
D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism; D055029:Inverted Repeat Sequences; D007700:Kinetics; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D043262:Ribonuclease P / Q000502:physiology*; D019562:Viral Load; D014779:Virus Replication
D017874:Immediate-Early Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; C048925:herpes simplex virus, type 1 protein ICP4; D043262:Ribonuclease P