IP Library › Granted Patent US 12,618,097
Granted Patent B2
US 12,618,097 · App. 17/240,009 · Granted May 5, 2026

Method for determining long non-coding ribonucleic acid interaction proteins

Inventors: Liang Zhang (New Territories, HK); Jian Yan (Kowloon, HK); Jingyu Li (Kowloon, HK); Wenkai Yi (New Territories, HK); Ligang Fan (Kowloon, HK)
Assignee: City University of Hong Kong
C12Q1/25C12N9/22C12N9/93C12N15/11G16B40/00C12N2310/20G01N2440/32
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Quick Facts
Patent No.
US 12,618,097
App. No.
17/240,009
Granted
May 5, 2026
Kind
B2
Abstract

The present invention provides a novel method for determining a long-chain non-coding ribonucleic acid interaction protein. The present invention provides a fusion protein formed by BASU and dCasRx, a mammalian expression vector for expressing said fusion protein. The method for determining the lncRNA interaction protein according to the present invention comprises: co-transfecting a mammalian expression vector that expresses the fusion protein and a gRNA that specifically targets the target lncRNA into target cells, thereby BASU specifically biotin-labeling effector proteins nearby; isolating the biotinylated proteins by using a streptavidin affinity coupled magnetic bead and then eluting, and digesting by trypsin and quantitatively analyzing by a label-free mass spectrometry. The present invention can highly credibly determine the proteins that interact with lncRNA.

Claims (10)

1 . A fusion protein formed by biotin ligase BASU and dCasRx, said fusion protein selected from the group consisting of B. subtilis biotin ligase with C-terminal mutation (BASU) as encoded by SEQ ID NO: 17-dCasRx as encoded by SEQ ID NO: 18 and dCasRx as encoded by SEQ ID NO: 18-BASU as encoded by SEQ ID NO: 17.

2 . A composition comprising:

the fusion protein according to claim 1 ; and

a pair of guide RNAs (gRNA) targeting a target long non-coding ribonucleic acid (lncRNA).

3 . A kit for determining lncRNA interaction proteins, comprising:

the fusion protein according to claim 1 ; and

a gRNA set targeting a target lncRNA.

4 . The composition according to claim 2 , wherein the target lncRNA is selected from the group consisting of X-inactivation specific transcript (XIST), Differentiation antagonizes non-protein-coding RNAs (DANCR), and Metastasis Associated Lung Adenocarcinoma Transcript 1 (MALAT1).

5 . The kit according to claim 3 , wherein the kit further comprises:

a control reagent without gRNA.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 27, 2021
From: ZHANG, LIANG; YAN, JIAN; LI, JINGYU; YI, WENKAI; FAN, LIGANG
To: CITY UNIVERSITY OF HONG KONG
Reel/Frame 056051/0569 →
Priority Claims (1)
CN 202010367970.0 · Apr 30, 2020 · national
Continuity (1)
Related Publication 20210340592A1 · Nov 4, 2021
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