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Allele specific targeting of APOE‐E4 expression: The novel CRISRP/CAS9‐based platform for precision therapy for Alzheimer’s disease
Author(s) -
Kantor Boris,
Moncalvo Malik,
Ilich Ekaterina,
Rittiner Joseph,
Dong Wendy,
Asmus Natalie,
Falek Carmel,
Yu Young Jun,
Gu Jeffrey,
Tringali Dominic,
ChibaFalek Ornit
Publication year - 2020
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1002/alz.036578
Subject(s) - dna methylation , biology , induced pluripotent stem cell , epigenetics , computational biology , epigenome , apolipoprotein e , gene , gene expression , genetics , disease , medicine , embryonic stem cell , pathology
Background APOEe4 is well‐established genetic risk factor for late onset Alzheimer’s disease (LOAD). It has been demonstrated that DNA methylation plays an important role in controlling the levels of APOEe4 expression. Furthermore, the differential methylation levels were reported between disease‐ and naïve‐ individuals. It also has been shown that reduction of APOEe4 levels may alleviate LOAD pathology. Here we aimed to develop a novel approach for specific and accurate modulation of APOEe4 gene expression. Method Here, we took advantage of a newly engineered VRER‐Cas9/gRNA system which specifically and accurately recognized a novel protospacer adjacent motif (PAM) created with SNP‐rs429358 T‐C transition in the APOEe4 gene. The system is based on an all‐in‐one lentiviral vector harboring gRNA/VRER‐dCas9‐DNMT3A. Using this vector, we were able to achieve an efficient enhancement in DNA methylation and the associated reduction in the expression of APOEe4 allele. The system has been validated on the isogeneic APOE‐human induced pluripotent stem cell (hiPSC)‐derived excitatory neurons. Result We were able to specifically and efficiently downregulate expression of APOEe4 using isogeneic APOE‐human induced pluripotent stem cell (hiPSC)‐derived excitatory neurons as the experimental model. The reduction in the gene expression was linked to robust and specific DNA methylation. Conclusion The developed epigenome‐editing platform highlights the novel approach towards the development of “smart” therapeutics for LOAD.