AAV8 Ins1-Cre can produce efficient β-cell recombination but requires consideration of off-target effects

In vivo genetic manipulation is used to study the impact of gene deletion or re-expression on β-cell function and organism physiology. Cre-LoxP is a system wherein LoxP sites flanking a gene are recognized by Cre recombinase. Cre transgenic mice are the most prevalent technology used to deliver Cre...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2020-06, Vol.10 (1), p.10518-10518, Article 10518
Main Authors: Ramzy, Adam, Tudurí, Eva, Glavas, Maria M, Baker, Robert K, Mojibian, Majid, Fox, Jessica K, O'Dwyer, Shannon M, Dai, Derek, Hu, Xiaoke, Denroche, Heather C, Edeer, Nazde, Gray, Sarah L, Verchere, Cameron B, Johnson, James D, Kieffer, Timothy J
Format: Article
Language:English
Subjects:
Animals
Beta cells
Cre recombinase
Dependovirus
Gene deletion
Hyperglycemia
Hypothalamus
Insulin
Insulin - genetics
Insulin - metabolism
Insulin-Secreting Cells - metabolism
Integrases
Mice
Mice, Transgenic
Pancreas
Promoter Regions, Genetic
Recombination
Recombination, Genetic
Rodents
Tamoxifen
Transgenic animals
Transgenic mice
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:In vivo genetic manipulation is used to study the impact of gene deletion or re-expression on β-cell function and organism physiology. Cre-LoxP is a system wherein LoxP sites flanking a gene are recognized by Cre recombinase. Cre transgenic mice are the most prevalent technology used to deliver Cre but many models have caveats of off-target recombination, impaired β-cell function, and high cost of animal production. Inducible estrogen receptor conjugated Cre models face leaky recombination and confounding effects of tamoxifen. As an alternative, we characterize an adeno associated virus (AAV) with a rat insulin 1 promoter driving Cre recombinase (AAV8 Ins1-Cre) that is economical and rapid to implement, and has limited caveats. Intraperitoneal AAV8 Ins1-Cre produced efficient β-cell recombination, alongside some hepatic, exocrine pancreas, α-cell, δ-cell, and hypothalamic recombination. Delivery of lower doses via the pancreatic duct retained good rates of β-cell recombination and limited rates of off-target recombination. Unlike inducible Cre in transgenic mice, AAV8 Ins1-Cre required no tamoxifen and premature recombination was avoided. We demonstrate the utility of this technology by inducing hyperglycemia in inducible insulin knockout mice (Ins1 ;Ins2 ). AAV-mediated expression of Cre in β-cells provides an effective alternative to transgenic approaches for inducible knockout studies.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-67136-w