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Novel cell phenotype screening method

A cell and phenotype technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of difficult gene expression reaction, mechanism analysis, slow speed, high cost, etc.

Pending Publication Date: 2022-07-15
新克赛特株式会社 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, in addition to the slow speed and high cost, it is difficult to quickly analyze the gene expression response, mechanism of action, etc. of individual cells in many ways.

Method used

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Examples

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Embodiment

[0219] Hereinafter, the present application will be specifically described based on examples, but the present application is not limited to these examples. In addition, unless otherwise specified, the measurement methods and units of this application conform to the Japanese Industrial Standards (JIS).

reference example 1

[0220] Reference Example 1: Preliminary test for the connection of the first barcoded nucleic acid to cells

[0221] According to the Multi-seq method (described in Nature Methods, Vol. 16, pp. 619-626, (2019)), the same cells, anchor CMOs, co-anchor CMOs and oligonucleotides were used as in Example 1 described below The acid was subjected to the following preliminary tests. That is, cells and anchor CMO were incubated in phosphate-buffered saline (PBS) solution at 4°C for 5 minutes, then co-anchor CMO was added thereto and further incubated at 4°C for 5 minutes, and finally, the red fluorescent A dye (Cy5) conjugated oligonucleotide (having a sequence corresponding to a partial sequence of the first barcode nucleic acid) was mixed with and incubated at 4°C for 5 minutes.

[0222] result as Figure 4 As shown in A and B in PBS solution, red fluorescent dye (Cy5)-conjugated oligonucleotides (with a sequence corresponding to the partial sequence of the first barcode nucleic ...

reference example 2

[0223] Reference Example 2: Preliminary test for the connection of the first barcoded nucleic acid to cells

[0224] Additionally, in addition to incubation with cell culture medium containing serum or bovine serum albumin (BSA) as solvent, use Test This preliminary experiment was carried out in the same manner as in Example 1. As a result, it was confirmed that the red fluorescent dye (Cy5)-conjugated oligonucleotide (having a sequence corresponding to the partial sequence of the first barcode nucleic acid) had a reduced rate of attachment to cells, such as Figure 4 Photos in C and D are shown.

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Abstract

The present application provides a method for screening cells, the method comprising: a step of preparing a plurality of cells labeled with a first barcode nucleic acid and treated with a test target, the first barcode nucleic acid relating to the test target; a step of selecting the plurality of cells based on cell phenotypes using an imaging cell sorter; and a step of identifying a test target for processing each cell using the first barcode nucleic acid as an indicator.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS [0002] This patent application claims priority based on US Patent Application Publication No. 62 / 959,420, filed January 10, 2020, and the entire disclosure of this prior patent application is hereby incorporated by reference into a part of the disclosure of this specification. technical field [0003] The present application relates to novel cellular phenotypic screening. Background technique [0004] Cell phenotyping (phenotypic screening) is widely known as a screening method for screening various drugs using cells. Phenotypic screening is the use of cell and organ phenotypes, such as cell proliferation rate, cell death, and cell image information as indicators of the localization of specific proteins or cellular structures, to find drugs that alter the phenotype of cells and organs (e.g., low molecular weight compounds, peptides, etc.). One of the important goals of cell phenotype screening is to examine the following inf...

Claims

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Application Information

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IPC IPC(8): C12Q1/6813C12Q1/04
CPCC12Q1/6806C12Q2563/107C12Q2563/159C12Q2563/179C12Q2600/106C12N15/1055C12N15/1065C12N15/1089C40B30/04
Inventor 坪内朝子太田祯生河崎史子
Owner 新克赛特株式会社
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