Improving virus production through quasispecies genomic selection and molecular breeding.
Virus production still is a challenging issue in antigen manufacture, particularly with slow-growing viruses. Deep-sequencing of genomic regions indicative of efficient replication may be used to identify high-fitness minority individuals suppressed by the ensemble of mutants in a virus quasispecies. Molecular breeding of quasispecies containing colonizer individuals, under regimes allowing more than one replicative cycle, is a strategy to select the fittest competitors among the colonizers. A slow-growing cell culture-adapted hepatitis A virus strain was employed as a model for this strategy. Using genomic selection in two regions predictive of efficient translation, the internal ribosome entry site and the VP1-coding region, high-fitness minority colonizer individuals were identified in a population adapted to conditions of artificially-induced cellular transcription shut-off. Molecular breeding of this population with a second one, also adapted to transcription shut-off and showing an overall colonizer phenotype, allowed the selection of a fast-growing population of great biotechnological potential.
Enteric Virus Laboratory, Department of Genetics, Microbiology and Statistics, School of Biology, University of Barcelona, Barcelona, Spain.|Enteric Virus Laboratory, Institute of Nutrition and Food Safety, Campus Torribera, University of Barcelona, Santa Coloma de Gramenet, Spain.
Francisco J
FJ
Pérez-Rodríguez
Enteric Virus Laboratory, Department of Genetics, Microbiology and Statistics, School of Biology, University of Barcelona, Barcelona, Spain.|Enteric Virus Laboratory, Institute of Nutrition and Food Safety, Campus Torribera, University of Barcelona, Santa Coloma de Gramenet, Spain.
Lucía
L
D'Andrea
Enteric Virus Laboratory, Department of Genetics, Microbiology and Statistics, School of Biology, University of Barcelona, Barcelona, Spain.|Enteric Virus Laboratory, Institute of Nutrition and Food Safety, Campus Torribera, University of Barcelona, Santa Coloma de Gramenet, Spain.
Montserrat
M
de Castellarnau
Enteric Virus Laboratory, Department of Genetics, Microbiology and Statistics, School of Biology, University of Barcelona, Barcelona, Spain.|Enteric Virus Laboratory, Institute of Nutrition and Food Safety, Campus Torribera, University of Barcelona, Santa Coloma de Gramenet, Spain.
Maria Isabel
MI
Costafreda
Enteric Virus Laboratory, Department of Genetics, Microbiology and Statistics, School of Biology, University of Barcelona, Barcelona, Spain.|Enteric Virus Laboratory, Institute of Nutrition and Food Safety, Campus Torribera, University of Barcelona, Santa Coloma de Gramenet, Spain.
Susana
S
Guix
Enteric Virus Laboratory, Department of Cell Biology, Physiology and Immunology, School of Biology, University of Barcelona, Barcelona, Spain.
Enric
E
Ribes
Liver Unit. Internal Medicine. Lab. Malalties Hepàtiques. Vall d'Hebron Institut Recerca-Hospital Universitari Vall d'Hebron (VHIR-HUVH), Barcelona, Spain.|Centro de Investigación Biomédica en Red (CIBER) de Enfermedades Hepáticas y Digestivas (CIBERehd) del Instituto de Salud Carlos III, Spain.
Josep
J
Quer
Liver Unit. Internal Medicine. Lab. Malalties Hepàtiques. Vall d'Hebron Institut Recerca-Hospital Universitari Vall d'Hebron (VHIR-HUVH), Barcelona, Spain.|Centro de Investigación Biomédica en Red (CIBER) de Enfermedades Hepáticas y Digestivas (CIBERehd) del Instituto de Salud Carlos III, Spain.|Roche Diagnostics SL, Barcelona, Spain.
Josep
J
Gregori
Enteric Virus Laboratory, Department of Genetics, Microbiology and Statistics, School of Biology, University of Barcelona, Barcelona, Spain.|Enteric Virus Laboratory, Institute of Nutrition and Food Safety, Campus Torribera, University of Barcelona, Santa Coloma de Gramenet, Spain.
Albert
A
Bosch
Enteric Virus Laboratory, Department of Genetics, Microbiology and Statistics, School of Biology, University of Barcelona, Barcelona, Spain.|Enteric Virus Laboratory, Institute of Nutrition and Food Safety, Campus Torribera, University of Barcelona, Santa Coloma de Gramenet, Spain.
Rosa M
RM
Pintó
D032621:DNA Shuffling; D016679:Genome, Viral; D023281:Genomics / Q000379:methods*; D006801:Humans; D010641:Phenotype; D000074704:Quasispecies / Q000235:genetics*; D014779:Virus Replication; D014780:Viruses / Q000235:genetics / Q000254:growth & development*