Somatic Editing of Ldlr With Adeno-Associated Viral-CRISPR Is an Efficient Tool for Atherosclerosis Research

Research output: Contribution to journalArticle

Kelsey E. Jarrett, Ciaran Lee, Marco De Giorgi, Ayrea Hurley, Baiba K. Gillard, Alexandria M. Doerfler, Ang Li, Henry J. Pownall, Gang Bao, William R. Lagor

Objective- Atherosclerosis studies in Ldlr knockout mice require breeding to homozygosity and congenic status on C57BL6/J background, a process that is both time and resource intensive. We aimed to develop a new method for generating atherosclerosis through somatic deletion of Ldlr in livers of adult mice. Approach and Results- Overexpression of PCSK9 (proprotein convertase subtilisin/kexin type 9) is currently used to study atherosclerosis, which promotes degradation of LDLR (low-density lipoprotein receptor) in the liver. We sought to determine whether CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats-associated 9) could also be used to generate atherosclerosis through genetic disruption of Ldlr in adult mice. We engineered adeno-associated viral (AAV) vectors expressing Staphylococcus aureus Cas9 and a guide RNA targeting the Ldlr gene (AAV-CRISPR). Both male and female mice received either (1) saline, (2) AAV-CRISPR, or (3) AAV-hPCSK9 (human PCSK9)-D374Y. A fourth group of germline Ldlr-KO mice was included for comparison. Mice were placed on a Western diet and followed for 20 weeks to assess plasma lipids, PCSK9 protein levels, atherosclerosis, and editing efficiency. Disruption of Ldlr with AAV-CRISPR was robust, resulting in severe hypercholesterolemia and atherosclerotic lesions in the aorta. AAV-hPCSK9 also produced hypercholesterolemia and atherosclerosis as expected. Notable sexual dimorphism was observed, wherein AAV-CRISPR was superior for Ldlr removal in male mice, while AAV-hPCSK9 was more effective in female mice. Conclusions- This all-in-one AAV-CRISPR vector targeting Ldlr is an effective and versatile tool to model atherosclerosis with a single injection and provides a useful alternative to the use of germline Ldlr-KO mice.

Original languageEnglish (US)
Pages (from-to)1997-2006
Number of pages10
JournalArteriosclerosis, Thrombosis, and Vascular Biology
Volume38
Issue number9
DOIs
StatePublished - Sep 1 2018

PMID: 30026278

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Somatic Editing of Ldlr With Adeno-Associated Viral-CRISPR Is an Efficient Tool for Atherosclerosis Research. / Jarrett, Kelsey E.; Lee, Ciaran; De Giorgi, Marco; Hurley, Ayrea; Gillard, Baiba K.; Doerfler, Alexandria M.; Li, Ang; Pownall, Henry J.; Bao, Gang; Lagor, William R.

In: Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 38, No. 9, 01.09.2018, p. 1997-2006.

Research output: Contribution to journalArticle

Harvard

Jarrett, KE, Lee, C, De Giorgi, M, Hurley, A, Gillard, BK, Doerfler, AM, Li, A, Pownall, HJ, Bao, G & Lagor, WR 2018, 'Somatic Editing of Ldlr With Adeno-Associated Viral-CRISPR Is an Efficient Tool for Atherosclerosis Research' Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 38, no. 9, pp. 1997-2006. https://doi.org/10.1161/ATVBAHA.118.311221

APA

Jarrett, K. E., Lee, C., De Giorgi, M., Hurley, A., Gillard, B. K., Doerfler, A. M., ... Lagor, W. R. (2018). Somatic Editing of Ldlr With Adeno-Associated Viral-CRISPR Is an Efficient Tool for Atherosclerosis Research. Arteriosclerosis, Thrombosis, and Vascular Biology, 38(9), 1997-2006. https://doi.org/10.1161/ATVBAHA.118.311221

Vancouver

Jarrett KE, Lee C, De Giorgi M, Hurley A, Gillard BK, Doerfler AM et al. Somatic Editing of Ldlr With Adeno-Associated Viral-CRISPR Is an Efficient Tool for Atherosclerosis Research. Arteriosclerosis, Thrombosis, and Vascular Biology. 2018 Sep 1;38(9):1997-2006. https://doi.org/10.1161/ATVBAHA.118.311221

Author

Jarrett, Kelsey E. ; Lee, Ciaran ; De Giorgi, Marco ; Hurley, Ayrea ; Gillard, Baiba K. ; Doerfler, Alexandria M. ; Li, Ang ; Pownall, Henry J. ; Bao, Gang ; Lagor, William R. / Somatic Editing of Ldlr With Adeno-Associated Viral-CRISPR Is an Efficient Tool for Atherosclerosis Research. In: Arteriosclerosis, Thrombosis, and Vascular Biology. 2018 ; Vol. 38, No. 9. pp. 1997-2006.

BibTeX

@article{a70de1ba3a894cb89a27d10cd5691c07,
title = "Somatic Editing of Ldlr With Adeno-Associated Viral-CRISPR Is an Efficient Tool for Atherosclerosis Research",
abstract = "Objective- Atherosclerosis studies in Ldlr knockout mice require breeding to homozygosity and congenic status on C57BL6/J background, a process that is both time and resource intensive. We aimed to develop a new method for generating atherosclerosis through somatic deletion of Ldlr in livers of adult mice. Approach and Results- Overexpression of PCSK9 (proprotein convertase subtilisin/kexin type 9) is currently used to study atherosclerosis, which promotes degradation of LDLR (low-density lipoprotein receptor) in the liver. We sought to determine whether CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats-associated 9) could also be used to generate atherosclerosis through genetic disruption of Ldlr in adult mice. We engineered adeno-associated viral (AAV) vectors expressing Staphylococcus aureus Cas9 and a guide RNA targeting the Ldlr gene (AAV-CRISPR). Both male and female mice received either (1) saline, (2) AAV-CRISPR, or (3) AAV-hPCSK9 (human PCSK9)-D374Y. A fourth group of germline Ldlr-KO mice was included for comparison. Mice were placed on a Western diet and followed for 20 weeks to assess plasma lipids, PCSK9 protein levels, atherosclerosis, and editing efficiency. Disruption of Ldlr with AAV-CRISPR was robust, resulting in severe hypercholesterolemia and atherosclerotic lesions in the aorta. AAV-hPCSK9 also produced hypercholesterolemia and atherosclerosis as expected. Notable sexual dimorphism was observed, wherein AAV-CRISPR was superior for Ldlr removal in male mice, while AAV-hPCSK9 was more effective in female mice. Conclusions- This all-in-one AAV-CRISPR vector targeting Ldlr is an effective and versatile tool to model atherosclerosis with a single injection and provides a useful alternative to the use of germline Ldlr-KO mice.",
keywords = "atherosclerosis, CRISPR-Cas systems, gene editing, hypercholesterolemia, lipoproteins",
author = "Jarrett, {Kelsey E.} and Ciaran Lee and {De Giorgi}, Marco and Ayrea Hurley and Gillard, {Baiba K.} and Doerfler, {Alexandria M.} and Ang Li and Pownall, {Henry J.} and Gang Bao and Lagor, {William R.}",
year = "2018",
month = "9",
day = "1",
doi = "10.1161/ATVBAHA.118.311221",
language = "English (US)",
volume = "38",
pages = "1997--2006",
journal = "Arteriosclerosis, thrombosis, and vascular biology",
issn = "1079-5642",
publisher = "Lippincott Williams and Wilkins",
number = "9",

}

RIS

TY - JOUR

T1 - Somatic Editing of Ldlr With Adeno-Associated Viral-CRISPR Is an Efficient Tool for Atherosclerosis Research

AU - Jarrett, Kelsey E.

AU - Lee, Ciaran

AU - De Giorgi, Marco

AU - Hurley, Ayrea

AU - Gillard, Baiba K.

AU - Doerfler, Alexandria M.

AU - Li, Ang

AU - Pownall, Henry J.

AU - Bao, Gang

AU - Lagor, William R.

PY - 2018/9/1

Y1 - 2018/9/1

N2 - Objective- Atherosclerosis studies in Ldlr knockout mice require breeding to homozygosity and congenic status on C57BL6/J background, a process that is both time and resource intensive. We aimed to develop a new method for generating atherosclerosis through somatic deletion of Ldlr in livers of adult mice. Approach and Results- Overexpression of PCSK9 (proprotein convertase subtilisin/kexin type 9) is currently used to study atherosclerosis, which promotes degradation of LDLR (low-density lipoprotein receptor) in the liver. We sought to determine whether CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats-associated 9) could also be used to generate atherosclerosis through genetic disruption of Ldlr in adult mice. We engineered adeno-associated viral (AAV) vectors expressing Staphylococcus aureus Cas9 and a guide RNA targeting the Ldlr gene (AAV-CRISPR). Both male and female mice received either (1) saline, (2) AAV-CRISPR, or (3) AAV-hPCSK9 (human PCSK9)-D374Y. A fourth group of germline Ldlr-KO mice was included for comparison. Mice were placed on a Western diet and followed for 20 weeks to assess plasma lipids, PCSK9 protein levels, atherosclerosis, and editing efficiency. Disruption of Ldlr with AAV-CRISPR was robust, resulting in severe hypercholesterolemia and atherosclerotic lesions in the aorta. AAV-hPCSK9 also produced hypercholesterolemia and atherosclerosis as expected. Notable sexual dimorphism was observed, wherein AAV-CRISPR was superior for Ldlr removal in male mice, while AAV-hPCSK9 was more effective in female mice. Conclusions- This all-in-one AAV-CRISPR vector targeting Ldlr is an effective and versatile tool to model atherosclerosis with a single injection and provides a useful alternative to the use of germline Ldlr-KO mice.

AB - Objective- Atherosclerosis studies in Ldlr knockout mice require breeding to homozygosity and congenic status on C57BL6/J background, a process that is both time and resource intensive. We aimed to develop a new method for generating atherosclerosis through somatic deletion of Ldlr in livers of adult mice. Approach and Results- Overexpression of PCSK9 (proprotein convertase subtilisin/kexin type 9) is currently used to study atherosclerosis, which promotes degradation of LDLR (low-density lipoprotein receptor) in the liver. We sought to determine whether CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats-associated 9) could also be used to generate atherosclerosis through genetic disruption of Ldlr in adult mice. We engineered adeno-associated viral (AAV) vectors expressing Staphylococcus aureus Cas9 and a guide RNA targeting the Ldlr gene (AAV-CRISPR). Both male and female mice received either (1) saline, (2) AAV-CRISPR, or (3) AAV-hPCSK9 (human PCSK9)-D374Y. A fourth group of germline Ldlr-KO mice was included for comparison. Mice were placed on a Western diet and followed for 20 weeks to assess plasma lipids, PCSK9 protein levels, atherosclerosis, and editing efficiency. Disruption of Ldlr with AAV-CRISPR was robust, resulting in severe hypercholesterolemia and atherosclerotic lesions in the aorta. AAV-hPCSK9 also produced hypercholesterolemia and atherosclerosis as expected. Notable sexual dimorphism was observed, wherein AAV-CRISPR was superior for Ldlr removal in male mice, while AAV-hPCSK9 was more effective in female mice. Conclusions- This all-in-one AAV-CRISPR vector targeting Ldlr is an effective and versatile tool to model atherosclerosis with a single injection and provides a useful alternative to the use of germline Ldlr-KO mice.

KW - atherosclerosis

KW - CRISPR-Cas systems

KW - gene editing

KW - hypercholesterolemia

KW - lipoproteins

UR - http://www.scopus.com/inward/record.url?scp=85053764444&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85053764444&partnerID=8YFLogxK

U2 - 10.1161/ATVBAHA.118.311221

DO - 10.1161/ATVBAHA.118.311221

M3 - Article

VL - 38

SP - 1997

EP - 2006

JO - Arteriosclerosis, thrombosis, and vascular biology

T2 - Arteriosclerosis, thrombosis, and vascular biology

JF - Arteriosclerosis, thrombosis, and vascular biology

SN - 1079-5642

IS - 9

ER -

ID: 42213948