ArticleJournal of clinical laboratory analysis2020
Utilizing CRISPR/Cas9 technology to prepare lymphoblastoid cell lines harboring genetic mutations for generating quality control materials in genetic testing.
Article in Journal of clinical laboratory analysis, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
3 citing papers in PubMed, 3 citations in OpenAlex.
- Development of a quality control panel based on patient-derived immortalized B-Lymphoblastoid cell lines for thalassemia genotyping.Molecular genetics and metabolism reports · 2026Article
- Development of novel quality control material based on CRISPR/Cas9 editing and xenografts for MLH1 protein deficiency testing.Journal of clinical laboratory analysis · 2021Article
- Utilizing CRISPR/Cas9 technology to prepare lymphoblastoid cell lines harboring genetic mutations for generating quality control materials in genetic testing.Journal of clinical laboratory analysis · 2020Article
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Authors and funding
9 authors at 1 institution in 1 country.
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Abstract
backgroundTo 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.
methodsIn 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 show evidence of predicted mutation.
resultsThe results showed that electroporation of the in vitro transcribed gRNAs into stable Cas9-expressing LCL was a more efficient gene-editing technique as compared to plasmid-mediated transfection, and that the positive rates could reach up to 35.9%. The predominance of indel sizes relative to the predicted deletion length was clustered between 10 and 0 bp. The results of whole-genome sequencing also demonstrated the existence of SEA-type deletion of α-thalassemia.
conclusionsGene-editing based on Cas9-expressing LCL by electroporation of sgRNA was a more efficient approach to introduce mutations for generating quality control materials for genetic testing. The edited lymphoblastoid cell lines were feasible to serve as quality control materials in genetic testing.
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