Comparative AAV-eGFP Transgene Expression Using Vector Serotypes 1-9, 7m8, and 8b in Human Pluripotent Stem Cells, RPEs, and Human and Rat Cortical Neurons

Stem Cells International
Thu T DuongJason A Mills

Abstract

Recombinant adeno-associated virus (rAAV), produced from a nonpathogenic parvovirus, has become an increasing popular vector for gene therapy applications in human clinical trials. However, transduction and transgene expression of rAAVs can differ across in vitro and ex vivo cellular transduction strategies. This study compared 11 rAAV serotypes, carrying one reporter transgene cassette containing a cytomegalovirus immediate-early enhancer (eCMV) and chicken beta actin (CBA) promoter driving the expression of an enhanced green-fluorescent protein (eGFP) gene, which was transduced into four different cell types: human iPSC, iPSC-derived RPE, iPSC-derived cortical, and dissociated embryonic day 18 rat cortical neurons. Each cell type was exposed to three multiplicity of infections (MOI: 1E4, 1E5, and 1E6 vg/cell). After 24, 48, 72, and 96 h posttransduction, GFP-expressing cells were examined and compared across dosage, time, and cell type. Retinal pigmented epithelium showed highest AAV-eGFP expression and iPSC cortical the lowest. At an MOI of 1E6 vg/cell, all serotypes show measurable levels of AAV-eGFP expression; moreover, AAV7m8 and AAV6 perform best across MOI and cell type. We conclude that serotype tropism is not only ca...Continue Reading

References

Apr 10, 1999·Journal of Virology·W XiaoJ M Wilson
Jan 29, 2003·Journal of Virology·Bernd Hauck, Weidong Xiao
Apr 29, 2008·The New England Journal of Medicine·Albert M MaguireJean Bennett
Jan 15, 2014·Alzheimer's & Dementia : the Journal of the Alzheimer's Association·Michael S RafiiRaymond T Bartus
Jun 5, 2014·Molecular Therapy : the Journal of the American Society of Gene Therapy·Yao LiStephen H Tsang
Jan 1, 2014·Molecular Therapy. Methods & Clinical Development·Nicolas CeresoVasiliki Kalatzis
Jun 9, 2015·Molecular Therapy. Methods & Clinical Development·Kleopatra RaptiRoger J Hajjar
Nov 6, 2015·Investigative Ophthalmology & Visual Science·Timothy A BlenkinsopJeffrey H Stern
Dec 1, 2017·Methods in Molecular Biology·Thilo M BuckJan Wijnholds
Dec 14, 2017·Molecular Therapy. Methods & Clinical Development·Gloriane SchnabolkBärbel Rohrer
Apr 21, 2018·Molecular Therapy : the Journal of the American Society of Gene Therapy·Ji Yun SongJean Bennett

❮ Previous
Next ❯

Citations

Nov 5, 2019·Frontiers in Cellular Neuroscience·Matthew V GreenStanley A Thayer
Feb 8, 2020·International Journal of Molecular Sciences·Marcela Garita-HernandezDeniz Dalkara
May 4, 2021·Frontiers in Cell and Developmental Biology·Marisa CappellaMaria Grazia Biferi

❮ Previous
Next ❯

Methods Mentioned

BETA
flow cytometry
electron microscopy
FACS
transfection

Software Mentioned

XLSTAT
ImageJ
Excel Analysis ToolPak
Accuri
matplotlib
Seaborn
AxioVision
excel
NIS Elements Advanced Research

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.