SaCas9 protein ssDNA aptamer SaCas9-04 and screening method and application thereof

The SaCas9-04 nucleic acid aptamer, prepared through a six-round screening method, solves the problem of insufficient specificity and affinity of SaCas9 protein ssDNA nucleic acid aptamers in existing technologies, and realizes efficient SaCas9 protein recognition and nucleic acid detection applications.

CN122235149APending Publication Date: 2026-06-19SHENZHEN SECOND PEOPLES HOSPITAL (SHENZHEN INST OF TRANSLATIONAL MEDICINE)
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
SHENZHEN SECOND PEOPLES HOSPITAL (SHENZHEN INST OF TRANSLATIONAL MEDICINE)
Filing Date
2026-05-19
Publication Date
2026-06-19

AI Technical Summary

Technical Problem

The lack of highly specific and affinity SaCas9 protein ssDNA aptamers in existing technologies limits its application potential in fields such as gene editing and nucleic acid detection.

Method used

Through a six-round screening method, the SaCas9 protein ssDNA aptamer SaCas9-04 was prepared. BSA and SaCas9 proteins were immobilized using carboxyl magnetic beads, and monoclonal nucleic acid aptamers with high binding capacity were screened by combining real-time PCR and high-throughput sequencing technology.

Benefits of technology

The obtained SaCas9-04 nucleic acid aptamer exhibits strong library binding ability, with a binding capacity greater than 30 RU and a dissociation constant KD of 739.2 pM. It possesses high specificity and stability and is suitable for the recognition, immobilization and nucleic acid detection of SaCas9 protein.

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Abstract

This invention relates to the field of biotechnology, and discloses a SaCas9 protein ssDNA aptamer, SaCas9-04, and its screening method and applications. The SaCas9-04 aptamer was obtained through six rounds of screening. Library affinity assays showed that the binding ability of the library increased with each screening round, with the sixth-round library exhibiting very strong affinity. Through analysis of high-throughput sequencing results, 96 single clones were selected for testing. Single-concentration screening using SPR was performed, successfully obtaining several single clones with binding values ​​greater than 30 RU, indicating good binding ability. Further concentration gradient KD testing of SaCas9-04 yielded a KD of 739.2 pM.
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