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Abstract / Description of output
The green alga Chlamydomonas reinhardtii is an invaluable reference organism to research fields including algal, plant, and ciliary biology. Accordingly, decades-long standing inefficiencies in targeted nuclear gene editing broadly hinder Chlamydomonas research. Here we report that single-step codelivery of CRISPR/Cpf1 ribonucleoproteins with single-stranded DNA repair templates results in precise and targeted DNA replacement with as much as ∼10% efficiency in C. reinhardtii. We demonstrate its use in transgene- and selection-free generation of sequence-specific mutations and epitope tagging at an endogenous locus. As the direct delivery of gene-editing reagents bypasses the use of transgenes, this method is potentially applicable to a wider range of species without the need to develop methods for stable transformation.
|Number of pages||6|
|Journal||Proceedings of the National Academy of Sciences (PNAS)|
|Early online date||5 Dec 2017|
|Publication status||Published - 19 Dec 2017|
Keywords / Materials (for Non-textual outputs)
- Chlamydomonas reinhardtii
FingerprintDive into the research topics of 'Efficient targeted DNA editing and replacement in Chlamydomonas reinhardtii using Cpf1 ribonucleoproteins and single-stranded DNA'. Together they form a unique fingerprint.
- 1 Finished
High value chemicals from algae: Developing a transgene-free genome editing toolbox to enhance carotenoid production
1/08/16 → 30/09/17