Table 3.
Neuronal target loci: gene expression.
| References | Locus* | Implication** | System | Delivery | Goal | Perturbation | Results |
|---|---|---|---|---|---|---|---|
| Amin et al., 2015 | miR-218-1; miR-218-2*** | Motoneuron disease | Mouse | Zygote injection of Cas9 mRNA and two sgRNAs | Germline transmission | KO; deletion | miR-218-1370bp−del or miR-218-2290bp−del mutant |
| Zuris et al., 2014 | NTF3 | Neurodegeneration | HEK293T | Cationic lipid transfection of (-30)GFP-TALE-VP64 protein or dCas9-VP64:sgRNA protein:RNA complex | Transient alteration of gene expression | – | Increased gene expression |
| Heller et al., 2014 | Fosb | Addiction, depression | Mouse (juvenile 7-8 wks old) | HSV transduction of ZF+effector (p65 or G9a) plasmids | Transient and local alteration of gene expression | Acetylation or methylation of promoter | Up or down-regulation of gene expression |
| HSV transduction of TALE+effector (VP64) plasmids | Transient and local alteration of gene expression | – | Increased gene expression | ||||
| Konermann et al., 2013 | Neurog2, Grm2+27 other neuronal genes | Neural differentiation, PD, Schizophrenia | N2a, mouse primary cortical neurons, mouse | Transfection of light-inducible TALE plasmids; transduction of light inducible AAV-TALE vectors | Light-induced transient alteration of gene expression | – | Increased gene expression with an improved temporal control |
| Maeder et al., 2013b | NTF3 | Neurodegeneration | HEK293 | dCas9-VP64 and sgRNA plasmid co-transfection | Transient alteration of gene expression | – | Increased gene expression |
| Chapdelaine et al., 2013 | FXN | Friedreich's ataxia | Human and mouse fibroblast | TALE-VP64 plasmid transfection | Transient alteration of gene expression | – | Increased gene expression |
| Maeder et al., 2013c | NTF3 | Neurodegeneration | HEK293 | TALE-VP64 or TALE-p65 plasmid transfection | Transient alteration of gene expression | – | Increased gene expression |
| Garriga-Canut et al., 2012 | HTT | HD | STHdh cells, mouse | ZFP plasmid or AAV2/1 vector transfection; | Transient or stable alteration of gene expression | – | Decreased gene expression |
| Cong et al., 2012 | CACNA1C | Autism | HEK293FT | TALE-VP64 plasmid transfection | Transient alteration of gene expression | – | Increased gene expression |
| Miller et al., 2011 | NTF3 | Neurodegeneration | HEK293 | TALE-VP64 plasmid transfection | Transient alteration of gene expression | – | Increased gene expression |
| Laganiere et al., 2010 | GDNF | PD | Human, monkey, and rat cells | Lentiviral transduction of ZF-p65 | Alteration of gene expression | – | Increased gene expression |
| Rat | AAV2 transduction of ZF-p65 | – |
Programmable DNA-binding agents (PDBAs; ZF, TALE, and RAN-guided Cas systems) are applied to modulate target gene expression.
Loci: CACNA1C, calcium channel voltage-dependent L-type alpha 1C subunit; Gdnf, glial cell line-derived neurotrophic factor, rat homolog of human GDNF; Grm2, glutamate receptor metabotropic 2, mouse homolog of human GRM2; HTT, huntingtin; miR-218-1, miR-218-2, microRNA-218 mammalian paralog 1, 2; Neurog2, neurogenin 2, mouse homolog of human NEUROG2; NTF3, neurotrophin 3.
Implication: HD, Huntington's disease; PD, Parkinson's disease.
miR-218 and miR-218-2 are knocked out through deletion (370 bp, 290 bp). This study by Amin et al. is included in gene expression modulation section because the indels in the regulatory elements mainly concern gene expression patterns.
(-30)GFP-TALE-VP64, engineered GFP protein with electrostatic charge of -30 fused to TALE-VP64; AAV2, adeno-associated virus type 2; Cas, CRISPR-associated; dCas, dead Cas; enzymatically inactive; G9a, histone methyltransferase, encoded by EHMT2, euchromatic histone-lysine N-methyltransferase 2; HA tag, human influenza hemagglutinin amino acids 98–106 epitope tag; HEK293, human embryonic kidney cell derivative line; HSV, herpes simplex virus; herpes simplex viral vector; indels, insertion and deletion mutations; KO, knock-out; N2a, mouse Neuro 2a cell line derived from neuroblastoma; p65, NF-κB activation domain; sgRNA, single guide RNA; STHdh, neuronal progenitor cell line from mouse embryonic day 14 striatal primordial, first exon of the mouse Htt gene has been replaced by a human HTT exon with 111 CAG repeats; TALE, transcription activator-like effector; TALENs, transcription activator-like effector nucleases; VP64, tetrameric repeat of herpes simplex virus activation domain (VP16); ZF, zinc finger; ZFN, zinc finger nuclease.