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

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.

Afiliación

Jarrett KE; From the Department of Molecular Physiology and Biophysics (K.E.J., M.D.G., A.H., A.M.D., W.R.L.).
Lee C; Integrative Molecular and Biomedical Sciences Graduate Program (K.E.J.), Baylor College of Medicine, Houston, TX.
De Giorgi M; Department of Bioengineering, Rice University, Houston, TX (C.L., A.L., G.B.).
Hurley A; From the Department of Molecular Physiology and Biophysics (K.E.J., M.D.G., A.H., A.M.D., W.R.L.).
Gillard BK; From the Department of Molecular Physiology and Biophysics (K.E.J., M.D.G., A.H., A.M.D., W.R.L.).
Doerfler AM; Houston Methodist Research Institute, TX (B.K.G., H.J.P.).
Li A; Weill Cornell Medicine, New York (B.K.G., H.J.P.).
Pownall HJ; From the Department of Molecular Physiology and Biophysics (K.E.J., M.D.G., A.H., A.M.D., W.R.L.).
Bao G; Department of Bioengineering, Rice University, Houston, TX (C.L., A.L., G.B.).
Lagor WR; Houston Methodist Research Institute, TX (B.K.G., H.J.P.).

Arterioscler Thromb Vasc Biol ; 38(9): 1997-2006, 2018 09.

Article en En | MEDLINE | ID: mdl-30026278

RESUMEN

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.

Asunto(s)

Aterosclerosis/genética; Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas; Modelos Animales de Enfermedad; Vectores Genéticos; Receptores de LDL/genética; Adenoviridae; Animales; Aterosclerosis/sangre; Proteína 9 Asociada a CRISPR/genética; Femenino; Edición Génica; Expresión Génica; Hipercolesterolemia/sangre; Hipercolesterolemia/genética; Hígado/metabolismo; Masculino; Ratones; Ratones Endogámicos C57BL; Ratones Noqueados; Proproteína Convertasa 9/sangre; Proproteína Convertasa 9/genética; Receptores de LDL/sangre

Palabras clave

CRISPR-Cas systems; atherosclerosis; gene editing; hypercholesterolemia; lipoproteins

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Receptores de LDL / Modelos Animales de Enfermedad / Aterosclerosis / Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas / Vectores Genéticos Tipo de estudio: Risk_factors_studies Límite: Animals Idioma: En Revista: Arterioscler Thromb Vasc Biol Asunto de la revista: ANGIOLOGIA Año: 2018 Tipo del documento: Article Pais de publicación: Estados Unidos

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google