Cell clones for cell phenotype research and their screening methods and applications

Abstract

Provided are a cell clone for cell phenotypic studies, a screening method therefor, and an application thereof. The screening method for the cell clone for cell phenotypic studies is mainly applicable when searching for a cell population of a certain cell phenotype in cells having not been subjected to experimental treatment. A gRNA barcode vector acts both as a barcode and a retrieval tool, thereby simplifying a searching and screening process. A first reporter gene of a gRNA-sensor search vector itself is not normally expressed, and signals only in response to action with a specific gRNA barcode and a nuclease for binding to the gRNA barcode, thereby allowing qualification in the search process. As long as a signal associated with the first reporter gene is present, the signal can be captured, thereby facilitating screening to obtain positive cell clones. The selected positive cells can be used in single studies, and are able to respond or generate phenotypes. The selected cells can also be widely used in studying cell phenotypes or in preparation of reagents for studying cell phenotypes.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a cell clone for cell phenotype research and a screening method and application thereof. Background technique [0002] In organisms, cells have different phenotypes, and it is very important to study the root causes of their different phenotypes. Taking cancer cells as an example, in cancer patients, tumor cell populations will show different phenotypes, such as some tumor cells can proliferate, some will migrate, and some are resistant to drugs, etc. These different phenotypes are the progress of cancer research. Therefore, understanding the basic biology of tumorigenesis is important to improve treatment outcomes. The fundamental reason why tumor cells have different phenotypes can be attributed to two mutually non-exclusive possibilities: one is that all tumor cells in a patient have the same intrinsic phenotypic potential, but there are many factors and variables in the pati...

AI Technical Summary

Problems solved by technology

However, there are many problems in traditional research methods, such as the use of shRNA interference carrier tools to retrieve barcodes, the process of reporter gene detection signals changing from strong to weak, and some cells are damaged due to changes in growth conditions and other external factors during the growth and passage process. It tends to weaken its own fluorescence, and it is easy to mix negative cells, which is not conducive to observation and screening; and if you want to further eliminate mixed cells, you need to use doxycycline to screen again, and the cells are treated again, which is complicated to operate. At the same time, the original characteristics of the cells may be affected; in addition, the retrieval process is a quantitative process, and it is not easy to retrieve those cells containing multiple copies of the barcode carrier

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