method

The parallel screening method using nucleotide constructs with PCR-based amplification addresses scalability and sequencing constraints in identifying cell-type-specific gene regulatory elements, enhancing the efficiency of AAV vector applications.

JP2026519978APending Publication Date: 2026-06-19SANIA RX LTD
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Patent Information

Authority / Receiving Office
JP · JP
Patent Type
Applications
Current Assignee / Owner
SANIA RX LTD
Filing Date
2024-05-13
Publication Date
2026-06-19

AI Technical Summary

Technical Problem

Existing methods for identifying cell-type-specific gene regulatory elements in AAV vectors are limited by scalability and practicality, as they require sequential testing in experimental animals and are constrained by the small packaging capacity and sequencing technology limitations.

Method used

A method for parallel screening of multiple putative gene regulatory elements using nucleotide constructs containing PRIMREG1-pGRE-PRIMREG2-GENE or GENE-PRIMREG1-pGRE-PRIMREG2 sequences, with PCR-based amplification of active and inactive pGREs, enabling high-speed, large-scale identification across cell and tissue types.

Benefits of technology

Enables scalable and efficient identification of cell-type-specific gene regulatory elements, reducing the need for sequential animal testing and overcoming sequencing limitations, thereby facilitating targeted gene therapies.

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Abstract

This disclosure relates to a novel screening method for screening multiple pGREs in parallel and to a nucleotide construct used in the method. The construct contains a nucleotide block PRIMREG1-pGRE-PRIMREG2-GENE or GENE-PRIMREG1-pGRE-PRIMREG2 in its sequence; where PRIMREG1 is either a primer recognition sequence 1 or an FLEx cassette containing primer recognition sequence 1; pGRE is a putative regulatory element; PRIMREG2 is either a primer recognition sequence 2 or an FLEx cassette containing primer recognition sequence 2; GENE contains nucleotide blocks REPSEQ, IRES and SSR; where REPSEQ is a reporter gene, which may or may not be present; IRES is either an internal ribosome entry site or a 2A autocleavage peptide; and SSR is a site-directed recombinase gene. At least one of PRIMREG1 and PRIMREG2 is an FLEx cassette containing a primer recognition sequence.
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