Invention Grant
- Patent Title: Activation of taste receptor genes in mammalian cells using CRISPR-Cas-9
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Application No.: US15762424Application Date: 2016-09-23
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Publication No.: US11459582B2Publication Date: 2022-10-04
- Inventor: Pul Ümit , Michael Krohn
- Applicant: B.R.A.I.N. AG
- Applicant Address: DE Zwingenberg
- Assignee: B.R.A.I.N. AG
- Current Assignee: B.R.A.I.N. AG
- Current Assignee Address: DE Zwingenberg
- Agency: Dilworth & Barrese, LLP
- Priority: EP15187005 20150926,EP15191360 20151024
- International Application: PCT/EP2016/072679 WO 20160923
- International Announcement: WO2017/050963 WO 20170330
- Main IPC: C12N15/90
- IPC: C12N15/90 ; C12N15/85 ; C12N9/22 ; C12N15/113 ; C12N15/63 ; C07K14/705 ; C12N15/11 ; C12N15/62 ; C07K19/00

Abstract:
Suggested is a method for enhancing the expression of taste related receptor genes encompassing the following steps: (i) providing a culture of mammalian cells, the genome of said cells comprising at least one sweet receptor domain; (ii) designing at least one type of single-guide RNA (sgRNA), the 10 to 30 nt guide sequence of said sgRNA being complementary to stretches within the non-coding and/or putative regulatory region upstream of the translation start codon of at least one sweet receptor gene; (iii) preparing a vector comprising an expression cassette encompassing at least one optionally modified CRISPR-Cas9, preferably CRISPR-dCas9VP64, and at least one optionally modified sg-RNA optionally containing aptamer structures for binding activator proteins; (iv) transfecting said culture of mammalian cells with said vector to target the genome for the presence of a DNA sequence that is complementary to the 10 to 30 nt guide sequence of said sgRNA; and (v) measuring the transcriptional enhancement of the sweet receptor mRNA by quantitative RT-PCR.
Public/Granted literature
- US20180273976A1 ACTIVATION OF TASTE RECEPTOR GENES IN MAMMALIAN CELLS USING CRISPR-CAS-9 Public/Granted day:2018-09-27
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