Site-directed Mutagenesis : Generation of an Extracistronic Mutation in Bacteriophage Qβ RNA

Trends in antiviral strategies

“…This was shown in the early site-directed mutagenesis experiments of bacteriophage Qb RNA performed by R. Flavell, C. Weissmann, and colleagues (Chapter 3). When the pyrimidine analog N 4 -hydroxy CMP was present at the extracistronic position 15, it directed the incorporation of GMP slightly more efficiently than AMP, while at position 39 incorporation of AMP was three-fold higher than GMP (compare Flavell et al, 1974 andDomingo et al, 1976). New mutagenic nucleotides are currently being investigated as potential lethal mutagens for viruses (Harki et al, 2002(Harki et al, , 2006(Harki et al, , 2007Graci and Cameron, 2004;Dapp et al, 2014;Vivet-Boudou et al, 2015; among other studies).…”

“…Favipiravir was discovered by Y. Furuta and colleagues as an effective antiviral agent against IV (Furuta et al, 2002(Furuta et al, , 2017. It has proven effective against a broad range of RNA viruses, with documented inhibitory, RNA chain termination and mutagenic activities in cell culture and in vivo [several studies are included in Table 9.1, and have been reviewed (Perales et al, 2019).…”

Reverse Genetics Meets the Nonsegmented Negative-Strand RNA Viruses

“…Especially useful is the application of reverse genetics which allows the genome of a virus (or an organism) to be changed at will by site-directed mutagenesis. The principle was established employing nucleotide analogues 1 in the case of the RNA bacteriophage Q ; today, mostly synthetic oligomers are used, as first demonstrated 2 with the DNA of the E. coli phage X174. Currently, incorporation of mutating oligonucleotides into cloned DNA is often carried out in a PCR.…”

Site-specific mutagenesis: retrospective and prospective

“…From the historical perspective, the framework for the genetic study of single lesions was established in 1974 by Weissmann and coworkers (30). They were able to introduce a single N 4 -hydroxy C into a predetermined site of the phage Q/3 RNA genome and, following its replication in vivo, showed that the progeny viral RNA contained U at this site.…”