Utilizing CRISPR/Cas9 technology to prepare lymphoblastoid cell lines harboring genetic mutations for generating quality control materials in genetic testing.

ABSTRACT:

Background

To meet the requirements of the rapidly progressing genetic testing technologies in clinical laboratories, assuring the quality of genetic tests by utilizing appropriate quality control materials is of paramount importance. The CRISPR/Cas9 technology was used to prepare quality control materials because genome-edited human cell lines are one of the major resources for quality control materials.

Methods

In this study, in vitro transcribed sgRNA were transfected into a Cas9-expressing lymphoblastoid cell line (LCL)-by electroporation-to simulate the SEA-type deletion observed in α-thalassemia. The edited positive cell line was screened and identified by polymerase chain reaction (PCR) followed by Sanger sequencing. The whole-genome sequencing was also performed to s

SUBMITTER: Zhou L 

PROVIDER: S-EPMC7370731 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature