A stable cell line carrying adenovirus-inducible rep and cap genes allows for infectivity titration of adeno-associated virus vectors.
Adeno-associated virus (AAV) vectors are being developed for in vivo and ex vivo gene transfer to human cells. At present, widespread usage of AAV vectors is limited primarily by difficulties in generating recombinant virions on a scale sufficient for in-depth preclinical and clinical trials. However, recent work in several laboratories suggests that this technical obstacle should be overcome in the near future. As a result, it can be anticipated that the interest in AAV vectors will expand, Thus, it becomes important to develop assay systems that will permit accurate quantification of the infectivity of AAV vectors derived from a variety of sources. We have developed an assay using a cell line that expresses AAV helper functions (rep and cap) upon induction by adenovirus infection. This assay system is based on the replication of input rAAV genomes rather than transgene expression (transduction). Thus, infectivity titrations in this system yield an estimation of rAAV infectious particles irrespective of the promoter or transgene present in the vector genome. Moreover, this assay method is more sensitive than conventional methods being used in other laboratories.
Clark
K R
KR
Department of Pediatrics, College of Medicine, Children's Hospital Research Foundation, Children's Hospital, Columbus, OH 43205, USA.
Voulgaropoulou
F
F
Johnson
P R
PR
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D002213:Capsid / Q000096:biosynthesis; D002460:Cell Line / Q000821:virology; D004268:DNA-Binding Proteins / Q000235:genetics; D000229:Dependovirus / Q000235:genetics / Q000302:isolation & purification*; D005814:Genes, Viral; D005822:Genetic Vectors / Q000235:genetics / Q000302:isolation & purification*; D006367:HeLa Cells; D006378:Helper Viruses / Q000235:genetics / Q000502:physiology; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D017186:Titrimetry; D014764:Viral Proteins / Q000235:genetics; D019065:Virus Assembly
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D014764:Viral Proteins; C068734:rep proteins, Adeno-associated virus 2