Repositorium
Efficient targeted DNA editing and replacement in Chlamydomonas reinhardtii using Cpf1 ribonucleoproteins and single-stranded DNA
419
Journal Article / 2017
Ferenczi, Aron; Pyott, Douglas Euan; Xipnitou, Andromachi; Molnar, Attila
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.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
967 |
Molnar, Attila United Kingdom |
Chlamydomonas reinhardtii |
CRISPR/Cpf1 FKB12 |
No information SDN1 |
Basic research Basic research |
968 |
Molnar, Attila United Kingdom |
Chlamydomonas reinhardtii |
CRISPR/Cpf1 FKB12 |
No information SDN2 |
Basic research Basic research |
969 |
Molnar, Attila United Kingdom |
Chlamydomonas reinhardtii |
CRISPR/Cpf1 FTSY |
bright green phenotype SDN2 |
Basic research Basic research |
970 |
Molnar, Attila United Kingdom |
Chlamydomonas reinhardtii |
CRISPR/Cpf1 SRP43 |
bright green phenotype SDN2 |
Basic research Basic research |
971 |
Molnar, Attila United Kingdom |
Chlamydomonas reinhardtii |
CRISPR/Cpf1 PHT7 |
No information SDN2 |
Basic research Basic research |
972 |
Molnar, Attila United Kingdom |
Nicotiana benthamiana |
CRISPR/Cpf1 GFP |
Green fluorescence SDN1 |
Basic research Basic research |