IP Library › Granted Patent US 12,285,495
Granted Patent B2
US 12,285,495 · App. 17/285,755 · Granted Apr 29, 2025

Genome editing methods and constructs

Inventors: Alberto Auricchio (Pozzuoli, IT); Manel Llado Santaeularia (Pozzuoli, IT)
Assignee: FONDAZIONE TELETHON ETS
A61K48/0058A61P27/02C12N9/22C12N15/11C12N15/907C12N2310/20C12N2750/14143
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Quick Facts
Patent No.
US 12,285,495
App. No.
17/285,755
Granted
Apr 29, 2025
Kind
B2
Abstract

The present invention relates to a method of integrating an exogenous DNA sequence into a genome of a cell comprising contacting the cell with: a) a donor nucleic acid comprising: —at least one STOP codon and a translation initiation sequence (TIS) or—a ribosomal skipping sequence, and—said exogenous DNA sequence wherein said donor nucleic acid is flanked at 5′ and 3′ by inverted targeting sequences; b) a complementary strand oligonucleotide homologous to the targeting sequence and c) a nuclease that recognizes the targeting sequence.

Claims (16)

1. A system comprising

a) a donor nucleic acid comprising

a STOP codon upstream to translation initiation sequence (TIS), and

an exogenous DNA sequence

wherein said donor nucleic acid is flanked at 5′ and 3′ by inverted targeting sequences, and wherein the TIS is a kozak consensus sequence comprising the sequence of SEQ ID NO: 54 or SEQ ID NO: 55 wherein n is g or a, or the TIS is a an IRES sequence comprising the sequence SEQ ID NO: 24 or SEQ ID NO: 23;

b) a complementary strand oligonucleotide homologous to the targeting sequence SEQ ID NO: 56-58 and

c) a nuclease that recognizes the targeting sequence.

2. An expression vector that comprises the system according to claim 1 .

3. The expression vector according to claim 2 , wherein the vector is selected from the group consisting of: Adena associated vector (AAV), adenoviral vector, lentiviral vector, retroviral vector or naked plasmid DNA vector.

4. A host cell comprising the system according to claim 1 .

5. A viral particle that comprises the system according to claim 1 .

6. The viral particles according to claim 5 , wherein the viral particle comprises capsid protein of an AAV.

7. The viral particle according to claim 6 , wherein the viral particle comprises capsid proteins of an AAV of a serotype selected from one or more of the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 AAV9 and AAV 10, preferably from the AAV2 or AAV8 serotype.

8. A pharmaceutical composition that comprises the system according to claim 1 and a pharmaceutical acceptable carrier.

9. A kit comprising: the system according to claim 1 in one or more containers.

10. The kit of claim 9 further comprising instructions or packaging materials that describe how to administer the system to a patient.

Assignments (2)
CHANGE OF NAME Recorded Nov 28, 2023
From: FONDAZIONE TELETHON
To: FONDAZIONE TELETHON ETS
Reel/Frame 065694/0135 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 11, 2022
From: AURICCHIO, ALBERTO; LLADO SANTAEULARIA, MANEL
To: FONDAZIONE TELETHON
Reel/Frame 061374/0095 →
Continuity (2)
Provisional Application 62745540 · Oct 15, 2018
Related Publication 20220001030A1 · Jan 6, 2022
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