The knock-in atlas: a web resource for targeted protein trap by CRISPR/Cas9 in human and mouse cell lines
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View abstract on PubMed
Summary
This summary is machine-generated.The Knock-in Atlas provides a valuable resource for creating gene knock-in cell lines using CRISPR-Cas9 technology. This web resource facilitates gene tagging with fluorescent proteins, aiding in cell engineering research.
Area Of Science
- Molecular Biology
- Genomics
- Cell Biology
Background
- CRISPR-Cas9 technology enables cell engineering, but large-scale targeted gene knock-in resources are limited.
- Efficient and reproducible gene knock-in is crucial for various biological studies.
Purpose Of The Study
- To introduce the Knock-in Atlas, a web resource for gene tagging via fluorescent protein knock-in.
- To provide a curated database of guide RNAs (gRNAs) and characterized knock-in cell lines.
Main Methods
- Developed knock-in cell lines by inserting artificial exons into target gene introns.
- Selected and cataloged guide RNAs (gRNAs) based on efficacy scores and protein structure.
- Characterized knock-in cell lines using flow cytometry, confocal microscopy, and western blotting.
Main Results
- Created knock-in cell lines for 350 proteins, with a focus on RNA binding proteins.
- Optimized transfection and flow cytometry protocols for multiple cell lines (HEK293T, eHAP1, HeLa, THP-1, Neuro2a, MEF, mESC).
- Launched a website (https://rnabio.naist.jp/atlas/) to share characterization data and pre-designed gRNA information.
Conclusions
- The Knock-in Atlas serves as a scalable resource for generating gene knock-in cell lines.
- This resource aids researchers in cell engineering and functional genomics studies.
- The curated gRNA database and characterized cell lines accelerate research in human and mouse genetics.
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