Details
- Autor(en) / Beteiligte
- Titel
- Heritable and precise zebrafish genome editing using a CRISPR-Cas system
- Ist Teil von
- PloS one, 2013-07, Vol.8 (7), p.e68708
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2013
- Link zum Volltext
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- We have previously reported a simple and customizable CRISPR (clustered regularly interspaced short palindromic repeats) RNA-guided Cas9 nuclease (RGN) system that can be used to efficiently and robustly introduce somatic indel mutations in endogenous zebrafish genes. Here we demonstrate that RGN-induced mutations are heritable, with efficiencies of germline transmission reaching as high as 100%. In addition, we extend the power of the RGN system by showing that these nucleases can be used with single-stranded oligodeoxynucleotides (ssODNs) to create precise intended sequence modifications, including single nucleotide substitutions. Finally, we describe and validate simple strategies that improve the targeting range of RGNs from 1 in every 128 basepairs (bps) of random DNA sequence to 1 in every 8 bps. Together, these advances expand the utility of the CRISPR-Cas system in the zebrafish beyond somatic indel formation to heritable and precise genome modifications.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0068708Titel-ID: cdi_plos_journals_1974583971
- Format
- –
- Schlagworte
- Animal genetics, Animals, Base pairs, Base Sequence, Biology, Cancer, CRISPR, CRISPR-Associated Proteins - chemistry, CRISPR-Associated Proteins - genetics, CRISPR-Cas Systems, Danio rerio, Deoxyribonucleic acid, DNA, DNA binding proteins, DNA methylation, DNA sequencing, Engineering, Genes, Genetic aspects, Genetic Engineering, Genome, Genomes, Genomics, Germ-Line Mutation, Hospitals, INDEL Mutation, Medical research, Medical schools, Medicine, Molecular Sequence Data, Mutagenesis, Mutagenesis, Insertional, Mutation, Mutation Rate, Nuclease, Nucleases, Nucleotide sequence, Nucleotide sequencing, Oligodeoxynucleotides, Oligonucleotides, Oligonucleotides - chemistry, Oligonucleotides - genetics, Pathology, Ribonucleic acid, RNA, Sequence Alignment, Zebrafish, Zebrafish - genetics