Promoter specific for non-pigmented ciliary epithelial cells
Inventors
Mahajan, Vinit • Wert, Katherine J.
Assignees
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Publication Number
US-12611469-B2
Publication Date
2026-04-28
Expiration Date
Abstract
A construct comprising a promoter specific for non-pigmented ciliary epithelial cells (NPCECs) is provided. In particular, the construct comprises a BEST2 minimal promoter that confers NPCEC-specific expression. The BEST2 minimal promoter may be operably linked to an expressible sequence such as a gene encoding a polypeptide of interest, a regulatory RNA sequence, a reporter gene, and the like. Such constructs may be provided in an expression vector, for example, with the BEST2 minimal promoter operably linked to an expressible sequence or in a host cell genetically modified with such an expression vector.
Core Innovation
The disclosed invention relates to a recombinant promoter construct that uses a minimal BEST2 promoter together with a transcriptional start site (TSS) to generate NPCEC-specific transcription. The promoter construct is transcriptionally active in non-pigmented ciliary epithelial cells (NPCECs) and not in retinal pigment epithelial cells (RPECs).
A key aspect is that the recombinant promoter construct has a non-native configuration, including non-native spacing between the minimal BEST2 promoter and the TSS. The minimal BEST2 promoter is defined by nucleotide sequence corresponding to positions 12,751,703-12,751,803 of human chromosome 19, or by the nucleotide sequence of SEO ID NO:1, or by a nucleotide sequence having at least 95% sequence identity to SEO ID NO:1, provided the promoter is capable of NPCEC-specific transcription.
The document further describes embodiments where the recombinant promoter construct is used in expression cassette and vector formats, including regulatory-element variants and selectable-marker vector formats. It also includes payload embodiments using expressible sequence, including polypeptides, regulatory RNA, and sequence-specific endonuclease systems such as CRISPR/Cas components, with the promoter/regulatory setup intended to drive expression selectively in NPCECs and not in RPECs.
Claims Coverage
The claims include one independent claim, which defines a recombinant promoter construct with NPCEC-specific activity and non-native promoter/TSS spacing. The dependent claims further refine the promoter by adding optional regulatory elements, specify payloads and expression cassette/vector embodiments, and include genome-editing and lentiviral structural elements, as explicitly described in the provided claim set.
Recombinant minimal BEST2 promoter with non-native spacing to TSS for NPCEC-specific transcription
A recombinant promoter construct comprising a minimal BEST2 promoter and a transcriptional start site (TSS), transcriptionally active in non-pigmented ciliary epithelial cells (NPCECs) and not in retinal pigment epithelial cells, with a non-native configuration including non-native spacing between the minimal BEST2 promoter and the TSS.
Minimal BEST2 promoter sequence definition for NPCEC-specific transcription
The minimal BEST2 promoter consists of (a) the nucleotide sequence corresponding to positions 12,751,703-12,751,803 of human chromosome 19, (b) the nucleotide sequence of SEO ID NO:1, or (c) a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEO ID NO:1, wherein the promoter is capable of NPCEC-specific transcription.
Overall, the claim coverage centers on promoter architecture: a minimal BEST2 promoter defined by specific sequence/identity criteria, combined with a TSS using non-native spacing to produce NPCEC-specific transcription and exclude transcription in retinal pigment epithelial cells. Dependent claim refinements add further regulatory elements and define downstream expression cassette/vector and payload classes, including genome-editing enzyme systems and lentiviral vector elements.
Stated Advantages
Provides NPCEC-specific transcription while not being active in retinal pigment epithelial cells.
Documented Applications
Local NPCEC-selective gene expression by introducing an expression vector into an eye, including preferential targeting after intravitreal injection as described in the document’s examples.
Using the promoter/regulatory setup to drive GFP expression selectively in NPCECs versus ARPE19, as demonstrated in example embodiments.
Using the promoter/regulatory setup for NPCEC-directed CRISPR/Cas9-mediated ADRB2 knockdown, as described in the document’s examples.
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