ArticleScientific reports2017
A small cassette enables conditional gene inactivation by CRISPR/Cas9.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 23 citations in OpenAlex.
- Universal conditional knockout approach to multiple species, from fish to human induced pluripotent stem cells.Nucleic acids research · 2026Article
- Ribonuclease 4 Functions in Nociceptor-Mediated Nerve Homeostasis.Nature communications · 2026Article
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- Evidence-Based Guide to Using Artificial Introns for Tissue-Specific Knockout in Mice.International journal of molecular sciences · 2023Review
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- Genetic and Molecular Quality Control of Genetically Engineered Mice.Methods in molecular biology (Clifton, N.J.) · 2023Article
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- Generation and Comparative Analysis of anNature cardiovascular research · 2022Article
- RNA methyltransferase METTL16: Targets and function.Wiley interdisciplinary reviews. RNA · 2022Review
- Designing and generating a mouse model: frequently asked questions.Journal of biomedical research · 2021Article
- ESI mutagenesis: a one-step method for introducing mutations into bacterial artificial chromosomes.Life science alliance · 2021Article
- Rapid Way to Generate Mouse Models forJournal of lipid and atherosclerosis · 2021Review
- Generation of mouse conditional knockout alleles in one step using theGenome research · 2021Article
- Pooled protein tagging, cellular imaging, and in situ sequencing for monitoring drug action in real time.Genome research · 2020Article
- Generation of Conditional Knockout Mice by Sequential Insertion of Two loxP Sites In Cis Using CRISPR/Cas9 and Single-Stranded DNA Oligonucleotides.Methods in molecular biology (Clifton, N.J.) · 2019Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The availability of CRISPR/Cas9 technology has enabled the rapid establishment of gene knockouts in many cell types and even whole organisms. However, conditional inactivation of essential genes remains a challenge. We devised an approach named DECAI (DEgradation based on Cre-regulated- Artificial Intron). It utilizes a small cassette of just 201 nucleotides that is inserted into the coding exon of a target gene using CRISPR/Cas9 technology and homology-directed repair. As its sequence is derived from an artificial intron, the cassette is removed by the splicing machinery and thus leaves no trace in the "off-state". Upon activation with Cre recombinase ("on-state"), the intron is crippled and the target gene is disrupted by a series of stop codons. We exemplify the utility of this approach on several non-essential and essential human genes. Clones bearing the conditional knockout cassette are recovered at frequencies above 5% and cassette function can be traced at the genomic DNA and the mRNA level. Importantly, cassette activation leads to loss of gene expression as judged by flow cytometry, Western blot or immunofluorescence. Altogether, this highlights the broad utility of the approach for conditional gene inactivation and suggests that this tool could be used to study the loss-of-function phenotypes of essential genes.
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