New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression in Retinal Ganglion Cells of Mouse and Primate Retina.
New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression in Retinal Ganglion Cells of Mouse and Primate Retina.
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DOI：
10.1089/hum.2018.118
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发表时间：
2019-03
影响因子：
4.2
通讯作者：
Wasserman WW
中科院分区：
文献类型：
--
作者：
Simpson EM;Korecki AJ;Fornes O;McGill TJ;Cueva-Vargas JL;Agostinone J;Farkas RA;Hickmott JW;Lam SL;Mathelier A;Renner LM;Stoddard J;Zhou M;Di Polo A;Neuringer M;Wasserman WW
Retinal gene therapy is leading the neurological gene therapy field, with 32 ongoing clinical trials of recombinant adeno-associated virus (rAAV)–based therapies. Importantly, over 50% of those trials are using restricted promoters from human genes. Promoters that restrict expression have demonstrated increased efficacy and can limit the therapeutic to the target cells thereby reducing unwanted off-target effects. Retinal ganglion cells are a critical target in ocular gene therapy; they are involved in common diseases such as glaucoma, rare diseases such as Leber's hereditary optic neuropathy, and in revolutionary optogenetic treatments. Here, we used computational biology and mined the human genome for the best genes from which to develop a novel minimal promoter element(s) designed for expression in restricted cell types (MiniPromoter) to improve the safety and efficacy of retinal ganglion cell gene therapy. Gene selection included the use of the first available droplet-based single-cell RNA sequencing (Drop-seq) dataset, and promoter design was bioinformatically driven and informed by a wide range of genomics datasets. We tested seven promoter designs from four genes in rAAV for specificity and quantified expression strength in retinal ganglion cells in mouse, and then the single best in nonhuman primate retina. Thus, we developed a new human-DNA MiniPromoter, Ple345 (NEFL), which in combination with intravitreal delivery in rAAV9 showed specific and robust expression in the retinal ganglion cells of the nonhuman-primate rhesus macaque retina. In mouse, we also developed MiniPromoters expressing in retinal ganglion cells, the hippocampus of the brain, a pan neuronal pattern in the brain, and peripheral nerves. As single-cell transcriptomics such as Drop-seq become available for other cell types, many new opportunities for additional novel restricted MiniPromoters will present.
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DOI：
10.1111/cts.12466
发表时间：
2017-07
期刊：
Clinical and translational science
影响因子：
--
作者：
Keeler AM;ElMallah MK;Flotte TR
通讯作者：
Flotte TR
影响因子：
4.3
作者：
Hanlon KS;Chadderton N;Palfi A;Blanco Fernandez A;Humphries P;Kenna PF;Millington-Ward S;Farrar GJ
通讯作者：
Farrar GJ
PAX6 MiniPromoters 在成年小鼠视网膜中驱动 rAAV 的限制性表达。
DOI：
10.1038/mtm.2016.51
发表时间：
2016
期刊：
Molecular therapy. Methods & clinical development
影响因子：
--
作者：
Hickmott JW;Chen CY;Arenillas DJ;Korecki AJ;Lam SL;Molday LL;Bonaguro RJ;Zhou M;Chou AY;Mathelier A;Boye SL;Hauswirth WW;Molday RS;Wasserman WW;Simpson EM
通讯作者：
Simpson EM
影响因子：
48
作者：
Danko CG;Hyland SL;Core LJ;Martins AL;Waters CT;Lee HW;Cheung VG;Kraus WL;Lis JT;Siepel A
通讯作者：
Siepel A
影响因子：
4.2
作者：
Flotte, Terence R.;Buening, Hildegard
通讯作者：
Buening, Hildegard