Highs and Lows in the Calicivirus Reverse Genetics DOI Open Access
Ángel L. Álvarez, Aroa Arboleya, Fábio A. Abade dos Santos

et al.

Published: March 26, 2024

In Virology, the term reverse genetics refers to a set of methodologies in which changes are introduced into viral genome and their effects on generation infectious progeny phenotypic features assessed. Reverse emerged thanks advances recombinant DNA technology made isolation, cloning, modification genes through mutagenesis possible. Most virus studies depend our capacity rescue an wild-type from cell cultures transfected with “infectious clone”. This clone generally consists circular plasmid containing functional copy full-length genome, under control appropriate RNA polymerase promoter. For most viruses, systems very straightforward, since genomes relatively easy handle modify also (with few notable exceptions) per se. is not true for whose need be transcribed cDNA before any -such as restriction enzyme digestion, gene knock-out or knock-in, site directed mutagenesis, ligation, etc.- can done. Establishing members Caliciviridae has proven exceptionally challenging, due low number this family that propagate cultures. Despite successful Feline calicivirus (FCV) more than 20 years ago, methods routine procedures easily extrapolated other caliciviruses. Reports have been far between, review, we discuss main pitfalls, failures, delays behind several clones.

Language: Английский

Highs and Lows in the Calicivirus Reverse Genetics DOI Open Access
Ángel L. Álvarez, Aroa Arboleya, Fábio A. Abade dos Santos

et al.

Published: March 26, 2024

In Virology, the term reverse genetics refers to a set of methodologies in which changes are introduced into viral genome and their effects on generation infectious progeny phenotypic features assessed. Reverse emerged thanks advances recombinant DNA technology made isolation, cloning, modification genes through mutagenesis possible. Most virus studies depend our capacity rescue an wild-type from cell cultures transfected with “infectious clone”. This clone generally consists circular plasmid containing functional copy full-length genome, under control appropriate RNA polymerase promoter. For most viruses, systems very straightforward, since genomes relatively easy handle modify also (with few notable exceptions) per se. is not true for whose need be transcribed cDNA before any -such as restriction enzyme digestion, gene knock-out or knock-in, site directed mutagenesis, ligation, etc.- can done. Establishing members Caliciviridae has proven exceptionally challenging, due low number this family that propagate cultures. Despite successful Feline calicivirus (FCV) more than 20 years ago, methods routine procedures easily extrapolated other caliciviruses. Reports have been far between, review, we discuss main pitfalls, failures, delays behind several clones.

Language: Английский

Citations

1