Literature summary for 3.1.26.5 extracted from

Bai, Y.; Li, H.; Vu, G.P.; Gong, H.; Umamoto, S.; Zhou, T.; Lu, S.; Liu, F.
Salmonella-mediated delivery of RNase P-based ribozymes for inhibition of viral gene expression and replication in human cells (2010), Proc. Natl. Acad. Sci. USA, 107, 7269-7274.

Application

Application	Comment	Organism
medicine	feasibility to develop Salmonella-mediated gene transfer of RNase P ribozymes as an effective approach for gene-targeting applications	synthetic construct

Cloned(Commentary)

Cloned (Comment)	Organism
DNA sequence coding for mutants M1-C1 and M1-C2 cloned into vector pU6, which contains a GFP expression cassette, and placed under the control of the small nuclear U6 RNA promoter. DNA sequence for ribozyme M1-thymidine kinase, which is derived from V57 and targets the herpes simplex virus 1 thymidine kinase mRNA, also cloned into vector pU6. Transformed into Salmonella typhimurium aroA strain SL7207	synthetic construct

Cloned (Comment)

Organism

DNA sequence coding for mutants M1-C1 and M1-C2 cloned into vector pU6, which contains a GFP expression cassette, and placed under the control of the small nuclear U6 RNA promoter. DNA sequence for ribozyme M1-thymidine kinase, which is derived from V57 and targets the herpes simplex virus 1 thymidine kinase mRNA, also cloned into vector pU6. Transformed into Salmonella typhimurium aroA strain SL7207

synthetic construct

Protein Variants

Protein Variants	Comment	Organism
A347C/C348U/C353G/C354GC355AG356U	M1-C2, is derived from M1-C1 by introducing several point mutations at the catalytic P4 domain. Is catalytically inactive	synthetic construct
G190U/A258C	functional ribozyme M1-C1, the 3'-terminus of an engineered M1GS ribozyme, V57, covalently linked with a guide sequence of 18 nucleotides that is complementary to the targeted mRNA sequence, which yields efficient cleavage	synthetic construct

Organism

Organism	UniProt	Comment	Textmining
synthetic construct	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	in vitro cleavage of human cytomegalovirus mRNA sequence by M1GS ribozyme. Incubation of a substrate containing the capsid scaffolding protein mRNA sequence with functional ribozyme M1-C1 (3'-terminus of an engineered M1GS ribozyme, V57, covalently linked with a guide sequence of 18 nucleotides that is complementary to the targeted mRNA sequence) yields efficient cleavage. Cleavage of capsid scaffolding protein mRNA by mutant M1-C2 or M1-thymidine kinase is barely detected	synthetic construct	?	-	?

Synonyms

Synonyms	Comment	Organism
M1GS	-	synthetic construct
ribonuclease P	-	synthetic construct
RNase P	-	synthetic construct

Expression

Organism	Comment	Expression
synthetic construct	Salmonella RNase P-based ribozyme sequence delivered in specific human cells, leads to substantial ribozyme expression. The M1GS RNAs appears to be exclusively expressed in the nuclei	up

General Information

General Information	Comment	Organism
malfunction	mutant M1-C2 is catalytically inactive	synthetic construct
physiological function	Salmonella can efficiently deliver RNase P-based ribozyme sequence in specific human cells, leading to substantial ribozyme expression and effective inhibition of viral infection: targeted gene delivery of RNase P ribozyme by Salmonella to human cytomegalovirus-infected cells results in effective inhibition of viral gene expression and replication. Functional RNase P ribozyme (M1GS RNA) that targets the overlapping mRNA region of two human cytomegalovirus capsid proteins, the capsid scaffolding protein and assemblin, which are essential for viral capsid formation. A reduction of 87-90% in viral capsid scaffolding protein expression and a reduction of about 5000fold in viral growth in cells that are treated with Salmonella carrying the sequence of the functional ribozyme	synthetic construct