A transient CRISPR/Cas9 expression system for genome editing in Trypanosoma brucei

Shaw, Sebastian; Knüsel, Sebastian; Hoenner, Sarah; Roditi, Isabel (2020). A transient CRISPR/Cas9 expression system for genome editing in Trypanosoma brucei. BMC research notes, 13(1), p. 268. Biomed Central 10.1186/s13104-020-05089-z

[img]
Preview
Text
s13104-020-05089-z.pdf - Published Version
Available under License Creative Commons: Attribution (CC-BY).

Download (1MB) | Preview

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.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Department of Biology > Institute of Cell Biology

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Shaw, Sebastian; Knüsel, Sebastian; Hoenner, Sarah Raphaela Maria and Roditi, Isabel

Subjects:

500 Science > 570 Life sciences; biology

ISSN:

1756-0500

Publisher:

Biomed Central

Funders:

[UNSPECIFIED] Swiss National Science Foundation ; [UNSPECIFIED] Canton of Bern

Language:

English

Submitter:

Isabel Roditi

Date Deposited:

15 Feb 2021 14:21

Last Modified:

21 Feb 2021 03:02

Publisher DOI:

10.1186/s13104-020-05089-z

PubMed ID:

32493474

BORIS DOI:

10.48350/152205

URI:

https://boris.unibe.ch/id/eprint/152205

Actions (login required)

Edit item Edit item
Provide Feedback