Activation of taste receptor genes in mammalian cells using CRISPR-Cas-9

US11459582B2 · kind B2 · utility

Assignee

• Brain Corporation · US

Inventors

• Pul Ümit · Duisburg, DE
• Michael Krohn · Lorsch, DE

Key dates

• Filing date: Sep 23, 2016
• Grant date: Oct 4, 2022
• Expiry date: Dec 24, 2037

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2830/00
• WIPO field: Biotechnology
• WIPO sector: Chemistry

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.