Preparation of multiplexed control materials for cancer mutation analysis by genome editing in GM12878 cells.

ABSTRACT:

Background

Quality control materials are necessary for assay development, test validation, and proficiency testing in cancer mutation analysis. Most of the existing controls for somatic mutations only harbor a single variant and are derived from unstable cell lines. This study aimed to establish a method to create stable multianalyte controls in a defined background by genome editing in GM12878 cells, which also can be applied for the reference of next-generation sequencing.

Methods

GM12878 cells were electroporated with a donor plasmid containing a mutant DNA sequence and a Cas9/sgRNA expressing vector. The genome-edited GM12878 cell was validated with Sanger sequencing, amplification refractory mutation system (ARMS), and next-generation sequencing (NGS).

Results

We

SUBMITTER: Lin G 

PROVIDER: S-EPMC8761438 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature