IP Library › Granted Patent US 12,553,063
Granted Patent B2
US 12,553,063 · App. 17/602,313 · Granted Feb 17, 2026

CAS13 family AAV vectors and uses thereof

Inventors: Guangping Gao (Worcester, MA); Alexander Brown (Worcester, MA)
Assignee: University of Massachusetts
C12N15/86A61P1/16C12N9/22C12N9/6424C12N15/113C12N15/907C12N2310/20C12N2330/51C12N2750/14143C12Y304/21
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Quick Facts
Patent No.
US 12,553,063
App. No.
17/602,313
Granted
Feb 17, 2026
Kind
B2
Abstract

Aspects of the disclosure relate to compositions and methods for multiplexed gene silencing in a cell or subject. In some embodiments, the disclosure provides an isolated nucleic acid or an rAAV encoding a transgene comprising a RNA-guided nuclease (RGN) operably linked to a first promoter, and a second promoter operably linked to a multi guide-RNA (multi-gRNA) expression cassette encoding one or more gRNAs targeting a gene associated with hypercholesterolemia or dyslipidemia. In some embodiments, the disclosure provides methods of treating a subject having hypercholesterolemia or dyslipidemia by administering the compositions.

Claims (27)

1 . An isolated nucleic acid encoding a transgene flanked by adeno-associated virus (AAV) inverted terminal repeats (ITRs), wherein the transgene comprises:

(i) a first nucleic acid sequence encoding an RNA-guided nuclease (RGN) comprising one or more sequences operably linked to a CB6 promoter; and

(ii) a second nucleic acid sequence comprising a multi guide-RNA (multi-gRNA) expression cassette encoding two or more gRNAs, each gRNA comprising the nucleotide sequence as set forth in any one of SEQ ID NOs: 1-56.

2 . The isolated nucleic acid of claim 1 , wherein the RGN is a Cas13 family protein or a variant thereof.

3 . The isolated nucleic acid of claim 1 , wherein the RGN is a Cas13RX or a dCas13 protein.

4 . The isolated nucleic acid of claim 1 , wherein the multi-gRNA cassette is operably linked to a second promoter.

5 . The isolated nucleic acid of claim 4 , wherein the second promoter is a U6 promoter.

6 . The isolated nucleic acid of claim 1 , wherein the multi-gRNA expression cassette encodes 3, 4, or 5 gRNAs.

7 . The isolated nucleic acid of claim 1 , wherein the multi-gRNA expression cassette comprises the following structure:

DR 30 -[guide sequence-DR 36 -guide sequence-DR 36 ] n -T

wherein:

DR 30 is a Cas13 direct repeat (DR) sequence comprising 30 nucleotides;

the guide sequence comprises between 10 and 35 nucleotides;

DR 36 is a Cas13 direct repeat (DR) sequence comprising 36 nucleotides; and

T is a terminator sequence; and

wherein n is an integer between 1 and 50.

8 . The isolated nucleic acid of claim 1 , wherein the transgene comprises one or more miRNA binding sites.

9 . The isolated nucleic acid of claim 1 , wherein at least one of the AAV ITRs is an AAV2 ITR.

10 . A recombinant adeno-associated virus (rAAV) comprising:

(i) the isolated nucleic acid of claim 1 ; and

(ii) a capsid protein.

11 . The rAAV of claim 10 , wherein the capsid protein has a tropism for liver tissue.

12 . The rAAV of claim 10 , wherein the capsid protein is an AAV8 or AAV9 capsid protein.

13 . The rAAV of claim 10 , wherein the rAAV is formulated for delivery to the liver.

14 . A composition comprising an isolated nucleic acid of claim 1 and a pharmaceutically acceptable excipient.

15 . A host cell comprising an isolated nucleic acid of claim 1 .

16 . The host cell of claim 15 , wherein the cell is a mammalian cell, bacterial cell, yeast cell, or insect cell.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 7, 2022
From: GAO, GUANGPING; BROWN, ALEXANDER
To: UNIVERSITY OF MASSACHUSETTS
Reel/Frame 059183/0950 →
Continuity (2)
Provisional Application 62833191 · Apr 12, 2019
Related Publication 20220186257A1 · Jun 16, 2022
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