ArticleFEBS open bio2014
Conditional targeting of Ispd using paired Cas9 nickase and a single DNA template in mice.
Article in FEBS open bio, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 38 citations in OpenAlex.
- Strategies and considerations for the generation of ssDNA-Based HDR templates for CRISPR-based genome editing.BMC genomics · 2026Review
- A nickase Cas9 gene-drive system promotes super-Mendelian inheritance in Drosophila.Cell reports · 2022Article
- Homology-based repair induced by CRISPR-Cas nucleases in mammalian embryo genome editing.Protein & cell · 2022Review
- CDP-ribitol prodrug treatment ameliorates ISPD-deficient muscular dystrophy mouse model.Nature communications · 2022Article
- Efficient and risk-reduced genome editing using double nicks enhanced by bacterial recombination factors in multiple species.Nucleic acids research · 2020Article
- Comparative analysis of single-stranded DNA donors to generate conditional null mouse alleles.BMC biology · 2018Article
- CDG Therapies: From Bench to Bedside.International journal of molecular sciences · 2018Review
- CRISPR/Cascade 9-Mediated Genome Editing-Challenges and Opportunities.Frontiers in genetics · 2018Review
- Easi-CRISPR for creating knock-in and conditional knockout mouse models using long ssDNA donors.Nature protocols · 2018Article
- Sexually dimorphic distribution of Prokr2 neurons revealed by the Prokr2-Cre mouse model.Brain structure & function · 2017Article
- CRISPR-Cas orthologues and variants: optimizing the repertoire, specificity and delivery of genome engineering tools.Mammalian genome : official journal of the International Mammalian Genome Society · 2017Review
- Mouse Genome Informatics (MGI) Resource: Genetic, Genomic, and Biological Knowledgebase for the Laboratory Mouse.ILAR journal · 2017Review
- Easi-CRISPR: a robust method for one-step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins.Genome biology · 2017Article
- Efficient gene targeting in mouse zygotes mediated by CRISPR/Cas9-protein.Transgenic research · 2017Article
- Optimization of the production of knock-in alleles by CRISPR/Cas9 microinjection into the mouse zygote.Scientific reports · 2017Article
- Exome sequencing in the knockin mice generated using the CRISPR/Cas system.Scientific reports · 2016Article
- Ultra-superovulation for the CRISPR-Cas9-mediated production of gene-knockout, single-amino-acid-substituted, and floxed mice.Biology open · 2016Article
- CRISPR/Cas9-Mediated Insertion of loxP Sites in the Mouse Dock7 Gene Provides an Effective Alternative to Use of Targeted Embryonic Stem Cells.G3 (Bethesda, Md.) · 2016Article
- Hyperlipidemia and hepatitis in liver-specific CREB3L3 knockout mice generated using a one-step CRISPR/Cas9 system.Scientific reports · 2016Article
- A CRISPR Path to Engineering New Genetic Mouse Models for Cardiovascular Research.Arteriosclerosis, thrombosis, and vascular biology · 2016Review
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
CRISPR/Cas9 technology is a highly promising genome editing tool in the mouse, potentially overcoming the costs and time required for more traditional gene targeting methods in embryonic stem (ES) cells. Recently, compared to the wildtype nuclease, paired Cas9 nickase (Cas9n) combined with single guide RNA (sgRNA) molecules has been found to enhance the specificity of genome editing while reducing off-target effects. Paired Cas9n has been shown to be as efficient as Cas9 for generating insertion and deletion (indel) mutations by non-homologous end joining and targeted deletion in the genome. However, an efficient and reliable approach to the insertion of loxP sites flanking critical exon(s) to create a conditional allele of a target gene remains an elusive goal. In this study, we microinjected Cas9n RNA with sgRNAs together with a single DNA template encoding two loxP sites flanking (floxing) exon 2 of the isoprenoid synthase containing domain (Ispd) into the pronucleus and cytoplasm of C57BL/6NCr one-cell stage zygotes. After surgical transfer, one F0 mouse expressing a conditional allele was produced (at a frequency of ∼8% of live pups born). The floxed allele was transmitted through the germline to F1 progeny, and could be successfully recombined using Cre recombinase. This study indicates that conditional targeting can be accomplished effectively using paired Cas9n and a single DNA template.
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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.