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Inhibition of herpes simplex virus 1 gene expression and replication by RNase P-associated external guide sequences. 2016

Jin Liu, and Luyao Shao, and Phong Trang, and Zhu Yang, and Michael Reeves, and Xu Sun, and Gia-Phong Vu, and Yu Wang, and Hongjian Li, and Congyi Zheng, and Sangwei Lu, and Fenyong Liu
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.

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.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D006367 HeLa Cells The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for, among other things, VIRUS CULTIVATION and PRECLINICAL DRUG EVALUATION assays. Cell, HeLa,Cells, HeLa,HeLa Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012333 RNA, Messenger RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm. Messenger RNA,Messenger RNA, Polyadenylated,Poly(A) Tail,Poly(A)+ RNA,Poly(A)+ mRNA,RNA, Messenger, Polyadenylated,RNA, Polyadenylated,mRNA,mRNA, Non-Polyadenylated,mRNA, Polyadenylated,Non-Polyadenylated mRNA,Poly(A) RNA,Polyadenylated mRNA,Non Polyadenylated mRNA,Polyadenylated Messenger RNA,Polyadenylated RNA,RNA, Polyadenylated Messenger,mRNA, Non Polyadenylated
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D014779 Virus Replication The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle. Viral Replication,Replication, Viral,Replication, Virus,Replications, Viral,Replications, Virus,Viral Replications,Virus Replications
D015870 Gene Expression The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION. Expression, Gene,Expressions, Gene,Gene Expressions
D015967 Gene Expression Regulation, Viral Any of the processes by which cytoplasmic factors influence the differential control of gene action in viruses. Regulation of Gene Expression, Viral,Viral Gene Expression Regulation,Regulation, Gene Expression, Viral
D043262 Ribonuclease P An RNA-containing enzyme that plays an essential role in tRNA processing by catalyzing the endonucleolytic cleavage of TRANSFER RNA precursors. It removes the extra 5'-nucleotides from tRNA precursors to generate mature tRNA molecules. Ribonuclease P, RNA Catalytic Subunit,RNA Catalytic Subunit, Ribonuclease P,RNase P
D055029 Inverted Repeat Sequences Copies of nucleic acid sequence that are arranged in opposing orientation. They may lie adjacent to each other (tandem) or be separated by some sequence that is not part of the repeat (hyphenated). They may be true palindromic repeats, i.e. read the same backwards as forward, or complementary which reads as the base complement in the opposite orientation. Complementary inverted repeats have the potential to form hairpin loop or stem-loop structures which results in cruciform structures (such as CRUCIFORM DNA) when the complementary inverted repeats occur in double stranded regions. Hairpin Loop Sequence,Inverted Repeat Sequence,Inverted Tandem Repeats,Palindromic Repeat Sequences,Sequence Palindromes,Stem-Loop Sequence,Hairpin Loop Sequences,Inverted Tandem Repeat,Palindrome, Sequence,Palindromes, Sequence,Palindromic Repeat Sequence,Repeat Sequence, Inverted,Repeat Sequence, Palindromic,Repeat Sequences, Inverted,Repeat Sequences, Palindromic,Repeat, Inverted Tandem,Repeats, Inverted Tandem,Sequence Palindrome,Sequence, Hairpin Loop,Sequence, Inverted Repeat,Sequence, Palindromic Repeat,Sequence, Stem-Loop,Sequences, Hairpin Loop,Sequences, Inverted Repeat,Sequences, Palindromic Repeat,Sequences, Stem-Loop,Stem Loop Sequence,Stem-Loop Sequences,Tandem Repeat, Inverted,Tandem Repeats, Inverted

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