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

Research output: Contribution to journalArticle

2 Citations (Scopus)
7 Downloads (Pure)

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.

Original languageEnglish
Pages (from-to)19-25
Number of pages7
JournalNeuropeptides
Volume64
Early online date3 Dec 2016
DOIs
Publication statusPublished - Aug 2017

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Neuropeptides
Animal Models
Neuropeptide Receptors
Genetically Modified Animals
Homologous Recombination
Appetite
Embryonic Stem Cells
Genes
Genome
Technology
Costs and Cost Analysis
Pain
Cell Line
Mutation
Health

Keywords

  • genome editing
  • non-homologous end-joining
  • homology-directed repair
  • off-target effects

Cite this

Using the CRISPR/Cas9 system to understand neuropeptide biology and regulation. / Hay, Elizabeth A; Knowles, Christopher; Kolb, Andreas; MacKenzie, Alasdair.

In: Neuropeptides, Vol. 64, 08.2017, p. 19-25.

Research output: Contribution to journalArticle

@article{b4d9497eabc745449dc4dad37013e9f5,
title = "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 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.",
keywords = "genome editing, non-homologous end-joining, homology-directed repair, off-target effects",
author = "Hay, {Elizabeth A} and Christopher Knowles and Andreas Kolb and Alasdair MacKenzie",
note = "Funding was provided by a Wellcome Trust ISSF starting grant (105625/Z/14/Z), Medical Research Scotland (PhD-719-2013), GW Pharmaceuticals (PhD-719-2013 - S.5242.001) and the BBSRC (BB/J012343/1).",
year = "2017",
month = "8",
doi = "10.1016/j.npep.2016.11.010",
language = "English",
volume = "64",
pages = "19--25",
journal = "Neuropeptides",
issn = "0143-4179",
publisher = "Churchill Livingstone",

}

TY - JOUR

T1 - Using the CRISPR/Cas9 system to understand neuropeptide biology and regulation

AU - Hay, Elizabeth A

AU - Knowles, Christopher

AU - Kolb, Andreas

AU - MacKenzie, Alasdair

N1 - Funding was provided by a Wellcome Trust ISSF starting grant (105625/Z/14/Z), Medical Research Scotland (PhD-719-2013), GW Pharmaceuticals (PhD-719-2013 - S.5242.001) and the BBSRC (BB/J012343/1).

PY - 2017/8

Y1 - 2017/8

N2 - 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.

AB - 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.

KW - genome editing

KW - non-homologous end-joining

KW - homology-directed repair

KW - off-target effects

U2 - 10.1016/j.npep.2016.11.010

DO - 10.1016/j.npep.2016.11.010

M3 - Article

VL - 64

SP - 19

EP - 25

JO - Neuropeptides

JF - Neuropeptides

SN - 0143-4179

ER -