ArticleeLife2017
Synthetically modified guide RNA and donor DNA are a versatile platform for CRISPR-Cas9 engineering.
Article in eLife, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 79 papers.
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Who cites it
79 citing papers in PubMed.
- Cyanine-modified ssODNs enhance CRISPR-Cas9 HDR in stem cell embryo models via chromatin and chemical modulation.Nature communications · 2026Article
- Advances in large-scale DNA engineering with the CRISPR system.Experimental & molecular medicine · 2025Review
- Improving RNA Secondary Structure Prediction Through Expanded Training Data.bioRxiv : the preprint server for biology · 2025Article
- Enhancing homology-directed repair efficiency with HDR-boosting modular ssDNA donor.Nature communications · 2024Article
- Near-perfect precise on-target editing of human hematopoietic stem and progenitor cells.eLife · 2024Article
- Recent advances in CRISPR-Cas9-based genome insertion technologies.Molecular therapy. Nucleic acids · 2024Review
- Self-delivering, chemically modified CRISPR RNAs for AAV co-delivery and genome editing in vivo.Nucleic acids research · 2024Article
- A novel GATA2 distal enhancer mutation results in MonoMAC syndrome in 2 second cousins.Blood advances · 2023Article
- Interstrand crosslinking of homologous repair template DNA enhances gene editing in human cells.Nature biotechnology · 2023Article
- Enrichment strategies to enhance genome editing.Journal of biomedical science · 2023Review
- Efficient site-specific integration in CHO-K1 cells using CRISPR/Cas9-modified donors.Molecular biology reports · 2023Article
- CATI: an efficient gene integration method for rodent and primate embryos by MMEJ suppression.Genome biology · 2023Article
- A Cell-Based Optimised Approach for Rapid and Efficient Gene Editing of Human Pluripotent Stem Cells.International journal of molecular sciences · 2023Article
- Strategies for precise gene edits in mammalian cells.Molecular therapy. Nucleic acids · 2023Review
- TAxI-peptide targeted Cas12a ribonuclease protein nanoformulations increase genome editing in hippocampal neurons.Journal of controlled release : official journal of the Controlled Release Society · 2023Article
- New advances in CRISPR/Cas-mediated precise gene-editing techniques.Disease models & mechanisms · 2023Review
- Online Databases of Genome Editing in Cardiovascular and Metabolic Diseases.Advances in experimental medicine and biology · 2023Article
- Practical Application of the 3Rs in Rodent Transgenesis.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Building Blocks of Artificial CRISPR-Based Systems beyond Nucleases.International journal of molecular sciences · 2022Review
- A Decade of CRISPR-Cas Gnome Editing inInternational journal of molecular sciences · 2022Review
19 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Chemical modification of the gRNA and donor DNA has great potential for improving the gene editing efficiency of Cas9 and Cpf1, but has not been investigated extensively. In this report, we demonstrate that the gRNAs of Cas9 and Cpf1, and donor DNA can be chemically modified at their terminal positions without losing activity. Moreover, we show that 5' fluorescently labeled donor DNA can be used as a marker to enrich HDR edited cells by a factor of two through cell sorting. In addition, we demonstrate that the gRNA and donor DNA can be directly conjugated together into one molecule, and show that this gRNA-donor DNA conjugate is three times better at transfecting cells and inducing HDR, with cationic polymers, than unconjugated gRNA and donor DNA. The tolerance of the gRNA and donor DNA to chemical modifications has the potential to enable new strategies for genome engineering.
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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.