title: Universal real-time PCR-based assay for lentiviral titration.
abstract: Lentiviral vectors are efficient vehicles for stable gene transfer in both dividing and non-dividing cells. This feature among others makes lentiviral vectors a powerful tool in molecular research. However, the use of lentiviruses in research studies and clinical trials requires a precise and validated titration method. In this study, we describe a qPCR-based approach for estimation of lentiviral vector titer (pLV-THM-GFP). The use of WPRE (Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element) and albumin genes as templates for an SYBR green-based real-time qPCR method allows for a rapid, sensitive, reproducible, and accurate assessment of lentiviral copy number at an integrated lentiviral DNA level. Furthermore, this optimization enables measurement of lentiviral concentration even in very poor quality and small quantity material. Consequently, this approach provides researchers with a tool to perform low-cost assessment with highly repeatable results.
authors.affiliation: Radiobiology Lab, Department of Medical Physics, The Greater Poland Cancer Centre, Garbary 15 Str., 61-866, Poznan, Poland, wojciech.barczak@wco.pl.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Barczak
authors.forename: Wiktoria
authors.initials: W
authors.lastname: Suchorska
authors.forename: Błażej
authors.initials: B
authors.lastname: Rubiś
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kulcenty
mesh_terms: D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000302:isolation & purification; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics* / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D049452:Green Fluorescent Proteins