ArticleScientific reports2018
High fidelity CRISPR/Cas9 increases precise monoallelic and biallelic editing events in primordial germ cells.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Who cites it
33 citing papers in PubMed, 69 citations in OpenAlex.
- From Design to Single-Cell Cloning: A Complete RNP-Based CRISPR-Cas9 Protocol for Precision Gene Correction in Human iPSCs.Stem cell reviews and reports · 2026Article
- A large animal model of heritable pulmonary arterial hypertension using BMPR2 gene-edited sheep.JCI insight · 2026Article
- Derivation of embryonic stem cells across avian species.Nature biotechnology · 2026Article
- Efficient and Safe Knockout ofVeterinary sciences · 2026Article
- Interleukin 10 controls the balance between tolerance, pathogen elimination, and immunopathology in birds.eLife · 2025Article
- Article
- Analysis of IDH1 and IDH2 mutations as causes of the hypermethylator phenotype in colorectal cancer.The Journal of pathology · 2025Article
- Tapping the microalgal potential: genetic precision and stress-induction for enhanced astaxanthin and biofuel production.Biotechnology for biofuels and bioproducts · 2025Review
- Application of CRISPR/Cas gene editing for infectious disease control in poultry.Open life sciences · 2025Review
- Harnessing the evolving CRISPR/Cas9 for precision oncology.Journal of translational medicine · 2024Review
- BUB1B monoallelic germline variants contribute to prostate cancer predisposition by triggering chromosomal instability.Journal of biomedical science · 2024Article
- Increasing Knockin Efficiency in Mouse Zygotes by Transient Hypothermia.The CRISPR journal · 2024Article
- Modeling mutation-specific arrhythmogenic phenotypes in isogenic human iPSC-derived cardiac tissues.Scientific reports · 2024Article
- Creating resistance to avian influenza infection through genome editing of the ANP32 gene family.Nature communications · 2023Article
- Enhancing Precision and Efficiency of Cas9-Mediated Knockin Through Combinatorial Fusions of DNA Repair Proteins.The CRISPR journal · 2023Article
- An optimized SpCas9 high-fidelity variant for direct protein delivery.Molecular therapy : the journal of the American Society of Gene Therapy · 2023Article
- CpG dinucleotide enrichment in the influenza A virus genome as a live attenuated vaccine development strategy.PLoS pathogens · 2023Article
- Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding.Plants (Basel, Switzerland) · 2023Review
- Development and function of chicken XCR1Frontiers in immunology · 2023Article
- Generation of Genome-Edited Chicken Through Targeting of Primordial Germ Cells.Methods in molecular biology (Clifton, N.J.) · 2023Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Primordial germ cells (PGCs), the embryonic precursors of the sperm and egg, are used for the introduction of genetic modifications into avian genome. Introduction of small defined sequences using genome editing has not been demonstrated in bird species. Here, we compared oligonucleotide-mediated HDR using wild type SpCas9 (SpCas9-WT) and high fidelity SpCas9-HF1 in PGCs and show that many loci in chicken PGCs can be precise edited using donors containing CRISPR/Cas9-blocking mutations positioned in the protospacer adjacent motif (PAM). However, targeting was more efficient using SpCas9-HF1 when mutations were introduced only into the gRNA target sequence. We subsequently employed an eGFP-to-BFP conversion assay, to directly compare HDR mediated by SpCas9-WT and SpCas9-HF1 and discovered that SpCas9-HF1 increases HDR while reducing INDEL formation. Furthermore, SpCas9-HF1 increases the frequency of single allele editing in comparison to SpCas9-WT. We used SpCas9-HF1 to demonstrate the introduction of monoallelic and biallelic point mutations into the FGF20 gene and generate clonal populations of edited PGCs with defined homozygous and heterozygous genotypes. Our results demonstrate the use of oligonucleotide donors and high fidelity CRISPR/Cas9 variants to perform precise genome editing with high efficiency in PGCs.
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Registered trials
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.