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Bio-Production of Adeno-Associated Virus for Gene Therapy

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Cell Culture Engineering and Technology

Part of the book series: Cell Engineering ((CEEN,volume 10))

Abstract

Genetic diseases are caused by absent or defective genes. Gene therapy aims to treat these conditions by delivering a functional copy of the affected gene into a patient’s cells. Gene delivery can be performed by recombinant viruses, such as Adeno-Associated Virus (AAV). AAVs are favoured due to their safety, low pathogenicity and their ability to infect multiple tissue types. Currently three AAV-based therapies are approved for the treatment of genetic diseases.

Genetic defects are usually present in every cell in a patient’s body. Therefore, gene therapy AAVs must be delivered to the majority of cells in an organ to achieve a therapeutic effect. As a result, AAV-based treatments require the administration of very high amounts of virus. Producing sufficient AAV, at an acceptable cost, without compromising safety is a major challenge in bio-production. Increasing or refining AAV production would enable the treatment of a wide variety of genetic diseases. Optimization strategies have focused on the genetic components needed for AAV assembly, the production cell lines, as well as omics-based approaches.

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Abbreviations

2DE :

2-dimensional gel electrophoresis

AAP :

Assembly activating protein

AAV :

Adeno-associated virus

AAVR :

Adeno-associated virus receptor

AAVS1 :

Adeno-associated virus integration site 1

BHK :

Baby hamster kidney cells

CFTR :

Cystic fibrosis transmembrane conductance regulator

E1/2/4 :

Adenovirus early region 1/2/4

FGFR :

Fibroblast growth factor receptor

FVIII :

Coagulation factor VIII

GeLC-MS :

Gel electrophoresis liquid chromatography-mass spectrometry

GFP :

Green fluorescent protein

GPN3 :

GPN-loop GTPase 3

GPR78 :

G protein-coupled receptor 78

HBoV1 :

Human bocavirus 1

HEK293 :

Human embryonic kidney 293 cells

HeLa :

Henrietta Lacks cells

HEM4 :

Uroporphyrinogen-III synthase

HGFR :

Hepatocyte growth factor receptor

HSV :

Herpes simplex viruses

ITR :

Inverted terminal repeat

LPLD :

Lipoprotein lipase deficiency

MAGv:

Multiplex AAV Genotyping

miRNA :

microRNA

ORF :

Open reading frame

PDGFR :

Platelet-derived growth factor receptor

PEI :

polyethyleneimine

PR E4 :

Proteinase yscE4

QC :

Quality control

qPCR :

quantitative polymerase chain reaction

rAAV :

recombinant Adeno-associated virus

RBE :

Rep binding element

Sf9/Sf21 :

Spodoptera frugiperda 9/21 cells

shRNA :

short hairpin RNA

SMN1 :

Survival motor neuron 1

SMRT :

Single molecule real-time

SOD1 :

Superoxide dismutase

TOP2 :

DNA topoisomerase 2

TRAP :

Tryptophan RNA-binding attenuation protein

trs :

Terminal resolution site

UTR :

Untranslated terminal region

VA-RNA :

Virus-associated RNA

VP1/2/3 :

Viral protein 1/2/3

ZF5 :

Zinc-finger 5 protein

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  1. National Institute of Bioprocessing, Research and Training, Dublin, Ireland

    Nicholas Donohue, Niamh Keogh, Stefano Boi & Niall Barron

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Donohue, N., Keogh, N., Boi, S., Barron, N. (2021). Bio-Production of Adeno-Associated Virus for Gene Therapy. In: Pörtner, R. (eds) Cell Culture Engineering and Technology. Cell Engineering, vol 10. Springer, Cham. https://doi.org/10.1007/978-3-030-79871-0_11

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