Journal article
A transient CRISPR/Cas9 expression system for genome editing in Trypanosoma brucei.
- Shaw S Institute of Cell Biology, University of Bern, Bern, Switzerland.
- Knüsel S Institute of Cell Biology, University of Bern, Bern, Switzerland.
- Hoenner S Institute of Cell Biology, University of Bern, Bern, Switzerland.
- Roditi I Institute of Cell Biology, University of Bern, Bern, Switzerland. isabel.roditi@izb.unibe.ch.
- 2020-06-05
Published in:
- BMC research notes. - 2020
English
OBJECTIVE
Generation of knockouts and in situ tagging of genes in Trypanosoma brucei has been greatly facilitated by using CRISPR/Cas9 as a genome editing tool. To date, this has entailed using a limited number of cell lines that are stably transformed to express Cas9 and T7 RNA polymerase (T7RNAP). It would be desirable, however, to be able to use CRISPR/Cas9 for any trypanosome cell line.
RESULTS
We describe a sequential transfection expression system that enables transient expression of the two proteins, followed by delivery of PCR products for gRNAs and repair templates. This procedure can be used for genome editing without the need for stable integration of the Cas9 and T7RNAP genes.
Generation of knockouts and in situ tagging of genes in Trypanosoma brucei has been greatly facilitated by using CRISPR/Cas9 as a genome editing tool. To date, this has entailed using a limited number of cell lines that are stably transformed to express Cas9 and T7 RNA polymerase (T7RNAP). It would be desirable, however, to be able to use CRISPR/Cas9 for any trypanosome cell line.
RESULTS
We describe a sequential transfection expression system that enables transient expression of the two proteins, followed by delivery of PCR products for gRNAs and repair templates. This procedure can be used for genome editing without the need for stable integration of the Cas9 and T7RNAP genes.
- Language
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- English
- Open access status
- gold
- Identifiers
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- DOI 10.1186/s13104-020-05089-z
- PMID 32493474
- Persistent URL
- https://sonar.ch/global/documents/178109
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