ArticleScientific reports2017
Optimization of the production of knock-in alleles by CRISPR/Cas9 microinjection into the mouse zygote.
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 36 papers.
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Who cites it
36 citing papers in PubMed, 76 citations in OpenAlex.
- Toward transgenic multicellular parasites: CRISPR-Cas12a germline editing ofScience advances · 2026Article
- Degron models: a toolbox for rapid in vivo depletion of essential proteins regulating mRNA metabolism.Communications biology · 2026Article
- Targeted Gene Knock-Out ofAnimals : an open access journal from MDPI · 2025Article
- The CRISPR-Cas9 System Is Used to Edit the Autoimmune Regulator Gene in Vitro and in Vivo.Advances in experimental medicine and biology · 2025Article
- Comprehensive posttranslational modifications in the testis-specific histone variant H3t protein validated in tagged knock-in mice.Scientific reports · 2024Article
- PCR-Based Strategy for Introducing CRISPR/Cas9 Machinery into Hematopoietic Cell Lines.Cancers · 2023Article
- Recent advances in the delivery and applications of nonviral CRISPR/Cas9 gene editing.Drug delivery and translational research · 2023Review
- Strategies for generation of mice via CRISPR/HDR-mediated knock-in.Molecular biology reports · 2023Review
- Innovative Strategies of Reprogramming Immune System Cells by Targeting CRISPR/Cas9-Based Genome-Editing Tools: A New Era of Cancer Management.International journal of nanomedicine · 2023Review
- CRISPR/Cas9-generated mouse model with humanizing single-base substitution in theFrontiers in genome editing · 2023Article
- CRISPR/Cas9: a tool to eradicate HIV-1.AIDS research and therapy · 2022Review
- CRISPR-Cas9-Based Technology and Its Relevance to Gene Editing in Parkinson's Disease.Pharmaceutics · 2022Review
- Introduction of loxP sites by electroporation in the mouse genome; a simple approach for conditional allele generation in complex targeting loci.BMC biotechnology · 2022Article
- Recent Advances in the Production of Genome-Edited Rats.International journal of molecular sciences · 2022Review
- Challenges of CRISPR-Based Gene Editing in Primary T Cells.International journal of molecular sciences · 2022Review
- Delivering the CRISPR/Cas9 system for engineering gene therapies: Recent cargo and delivery approaches for clinical translation.Frontiers in bioengineering and biotechnology · 2022Review
- Applications of and considerations for using CRISPR-Cas9-mediated gene conversion systems in rodents.Nature protocols · 2022Review
- Modification of improved-genome editing via oviductal nucleic acids delivery (i-GONAD)-mediated knock-in in rats.BMC biotechnology · 2021Article
- Exosome/Liposome-like Nanoparticles: New Carriers for CRISPR Genome Editing in Plants.International journal of molecular sciences · 2021Review
- Early onset severe ATP1A2 epileptic encephalopathy: Clinical characteristics and underlying mutations.Epilepsy & behavior : E&B · 2021Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Microinjection of the CRISPR/Cas9 system in zygotes is an efficient and comparatively fast method to generate genetically modified mice. So far, only few knock-in mice have been generated using this approach, and because no systematic study has been performed, parameters controlling the efficacy of CRISPR/Cas9-mediated targeted insertion are not fully established. Here, we evaluated the effect of several parameters on knock-in efficiency changing only one variable at a time. We found that knock-in efficiency was dependent on injected Cas9 mRNA and single-guide RNA concentrations and that cytoplasmic injection resulted in more genotypic complexity compared to pronuclear injection. Our results also indicated that injection into the pronucleus compared to the cytoplasm is preferable to generate knock-in alleles with an oligonucleotide or a circular plasmid. Finally, we showed that Cas9D10A nickase variant was less efficient than wild-type Cas9 for generating knock-in alleles and caused a higher rate of mosaicism. Thus, our study provides valuable information that will help to improve the future production of precise genetic modifications in mice.
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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.