Methods and compositions for donating DNA molecules

By using circular single-stranded DNA (cssDNA) donor molecules to form a double-stranded DNA region with substrate complementary oligonucleotides (SCO), and then combining with endogenous DNA polymerase for second-strand synthesis, the risks of immune sensing and integration of dsDNA donor molecules in the cell nucleus are resolved, enabling safe and efficient genome insertion.

CN122374459APending Publication Date: 2026-07-10THE GENERAL HOSPITAL CORP
10 Cites 0 Cited by

Patent Information

Application Number
CN202480078465.0
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Priority Date
2024-04-24
Filing Date
2024-10-11
Publication Date
2026-07-10

AI Technical Summary

Technical Problem

In the prior art, double-stranded DNA (dsDNA) donor molecules have undesirable off-target integration and stimulation of cellular immune response when delivered to the cell nucleus. Furthermore, traditional DNA target enzymes recognize double-stranded DNA rather than single-stranded DNA, leading to risks of immune response and integration.

Method used

Using a circular single-stranded DNA (cssDNA) donor molecule, a substrate complementary oligonucleotide (SCO) is bound to form a double-stranded DNA region containing a recombinase or transposase recognition sequence. The second strand is then synthesized in the cell nucleus by an endogenous DNA polymerase, thus achieving integration.

Benefits of technology

It reduces the risk of cellular immune induction, improves the delivery efficiency and safety of DNA donor molecules, and enables efficient genome insertion.

✦ Generated by Eureka AI based on patent content.
Patent Text Reader

Abstract

We describe compositions of DNA donor molecules compatible with various gene insertion methods that have major advantages in delivery, nuclear localization and immune escape. The modified DNA donor molecule is predominantly single-stranded, but contains a short region of double-stranded DNA to reconstruct the enzyme binding site, and it can be combined with other methods to enhance donor recruitment to the nucleus.
Need to check novelty before this filing date? Find Prior Art