Using the CRISPR/Cas9 system to understand neuropeptide biology and regulation

Abstract Neuropeptides and their receptors play a role in physiological responses such as appetite, stress and inflammatory pain. With neuropeptides having such diverse and important physiological roles, knocking-out the genes encoding them, their receptors, parts of their regulatory sequences, or r...

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Bibliographic Details
Published in:Neuropeptides (Edinburgh) 2017-08, Vol.64, p.19-25
Main Authors: Hay, Elizabeth A, Knowles, Christopher, Kolb, Andreas, MacKenzie, Alasdair
Format: Article
Language:English
Subjects:
Advanced Basic Science
Animal models
Animals
Cell lines
Cellular biology
CRISPR
CRISPR-Cas Systems - genetics
Embryo cells
Endocrinology & Metabolism
Gene Expression - genetics
Gene Knock-In Techniques - methods
Gene Knockout Techniques - methods
Gene regulation
Genome editing
Genomes
Homologous recombination
Homology-directed repair
Humans
Inflammation
Mutation
Mutation - genetics
Neuropeptides
Neuropeptides - genetics
Non-homologous end-joining
Off-target effects
Pain
Regulatory sequences
Stem cell transplantation
Stem cells
Transgenic animals
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Summary:Abstract Neuropeptides and their receptors play a role in physiological responses such as appetite, stress and inflammatory pain. With neuropeptides having such diverse and important physiological roles, knocking-out the genes encoding them, their receptors, parts of their regulatory sequences, or reproducing disease associated polymorphic variants are important steps in studying neuropeptides and how they may contribute to disease. Previously, knock-outs were generated using methods such as targeted homologous recombination in embryonic stem cells but this method is costly and time-consuming. The CRISPR/Cas9 system has rapidly taken over the genome editing field and will advance our understanding of neuropeptide genes and their regulation. With CRISPR/Cas9 technology, the time and costs involved in producing transgenic animal models, is greatly reduced. In this review, we describe how the system can be used to manipulate genomic sequences by “knock-out” or “knock-in” mutations in cell lines or in animal models. We also discuss the specificity of the system and methods to limit off-target effects. When combined with the availability of genome sequences, CRISPR/Cas9 directed genome editing in vitro and in vivo, promises to provide a deeper understanding of the biology of the neuropeptides in health and disease than has ever been available before.
ISSN:0143-4179
1532-2785
DOI:10.1016/j.npep.2016.11.010