ArticleRNA (New York, N.Y.)2019
A piggyBac-based toolkit for inducible genome editing in mammalian cells.
Article in RNA (New York, N.Y.), 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 45 citations in OpenAlex.
- Review
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- Structural and systems characterization of phosphorylation on metabolic enzymes identifies sex-specific metabolic reprogramming in obesity.Molecular cell · 2025Article
- Unraveling the cohesin-chromatin interface: identifying protein interactions that modulate chromosome structure and function.Epigenetics & chromatin · 2025Article
- Isogenic comparison of Airn and Xist reveals core principles of Polycomb recruitment by lncRNAs.Molecular cell · 2025Article
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- XIST dampens X chromosome activity in a SPEN-dependent manner during early human development.Nature structural & molecular biology · 2024Article
- The interferon γ pathway enhances pluripotency and X-chromosome reactivation in iPSC reprogramming.Science advances · 2024Article
- Precise large-fragment deletions in mammalian cells and mice generated by dCas9-controlled CRISPR/Cas3.Science advances · 2024Article
- Parallelized engineering of mutational models using piggyBac transposon delivery of CRISPR libraries.Cell reports methods · 2024Article
- SAFB associates with nascent RNAs and can promote gene expression in mouse embryonic stem cells.RNA (New York, N.Y.) · 2023Article
- Rho MultiBinder, a fluorescent biosensor that reports the activity of multiple GTPases.Biophysical journal · 2023Article
- Proximity-dependent recruitment of Polycomb repressive complexes by the lncRNA Airn.Cell reports · 2023Article
- H4K16ac activates the transcription of transposable elements and contributes to their cis-regulatory function.Nature structural & molecular biology · 2023Article
- A monoclonal antibody raised against human EZH2 cross-reacts with the RNA-binding protein SAFB.Biology open · 2023Article
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
Abstract
We describe the development and application of a novel series of vectors that facilitate CRISPR-Cas9-mediated genome editing in mammalian cells, which we call CRISPR-Bac. CRISPR-Bac leverages the piggyBac transposon to randomly insert CRISPR-Cas9 components into mammalian genomes. In CRISPR-Bac, a single piggyBac cargo vector containing a doxycycline-inducible Cas9 or catalytically dead Cas9 (dCas9) variant and a gene conferring resistance to Hygromycin B is cotransfected with a plasmid expressing the piggyBac transposase. A second cargo vector, expressing a single-guide RNA (sgRNA) of interest, the reverse-tetracycline TransActivator (rtTA), and a gene conferring resistance to G418, is also cotransfected. Subsequent selection on Hygromycin B and G418 generates polyclonal cell populations that stably express Cas9, rtTA, and the sgRNA(s) of interest. We show that CRISPR-Bac can be used to knock down proteins of interest, to create targeted genetic deletions with high efficiency, and to activate or repress transcription of protein-coding genes and an imprinted long noncoding RNA. The ratio of sgRNA-to-Cas9-to-transposase can be adjusted in transfections to alter the average number of cargo insertions into the genome. sgRNAs targeting multiple genes can be inserted in a single transfection. CRISPR-Bac is a versatile platform for genome editing that simplifies the generation of mammalian cells that stably express the CRISPR-Cas9 machinery.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.