Rapid Testing of CRISPR Nucleases and Guide RNAs in an Cell-Free Transcription-Translation System.
Name:
Publisher version
View Source
Access full-text PDFOpen Access
View Source
Check access options
Check access options
Name:
Marshall, Beisel and Noireau.pdf
Size:
822.7Kb
Format:
PDF
Description:
Open Access publication
Average rating
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Star rating
Your vote was cast
Thank you for your feedback
Thank you for your feedback
Issue Date
2020-06-03
Metadata
Show full item recordAbstract
We present a protocol to rapidly test DNA binding and cleavage activity by CRISPR nucleases using cell-free transcription-translation (TXTL). Nuclease activity is assessed by adding DNA encoding a nuclease, a guide RNA, and a targeted reporter to a TXTL reaction and by measuring the fluorescence for several h. The reactions, performed in a few microliters, allow for parallel testing of many nucleases and guide RNAs. The protocol includes representative results for (d)Cas9 from Streptococcus pyogenes targeting a GFP reporter gene. For complete information on the generation and use of this protocol, please refer to the paper by Marshall et al. (2018).Citation
STAR Protoc. 2020 Jun 3;1(1):100003. doi: 10.1016/j.xpro.2019.100003.Affiliation
HIRI, Helmholtz-Institut für RNA-basierte Infektionsforschung, Josef-Shneider Strasse 2, 97080 Würzburg, Germany.Publisher
Elsevier (CellPress)Journal
STAR protocolsPubMed ID
33111065Type
ArticleLanguage
enEISSN
2666-1667ae974a485f413a2113503eed53cd6c53
10.1016/j.xpro.2019.100003
Scopus Count
The following license files are associated with this item:
- Creative Commons
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-ShareAlike 4.0 International
Related articles
- Rapid and Scalable Characterization of CRISPR Technologies Using an E. coli Cell-Free Transcription-Translation System.
- Authors: Marshall R, Maxwell CS, Collins SP, Jacobsen T, Luo ML, Begemann MB, Gray BN, January E, Singer A, He Y, Beisel CL, Noireaux V
- Issue date: 2018 Jan 4
- Rapidly Characterizing CRISPR-Cas13 Nucleases Using Cell-Free Transcription-Translation Systems.
- Authors: Wandera KG, Beisel CL
- Issue date: 2022
- A detailed cell-free transcription-translation-based assay to decipher CRISPR protospacer-adjacent motifs.
- Authors: Maxwell CS, Jacobsen T, Marshall R, Noireaux V, Beisel CL
- Issue date: 2018 Jul 1
- Targeted genome editing in human cells using CRISPR/Cas nucleases and truncated guide RNAs.
- Authors: Fu Y, Reyon D, Joung JK
- Issue date: 2014
- Nucleic Acid-Dependent Conformational Changes in CRISPR-Cas9 Revealed by Site-Directed Spin Labeling.
- Authors: Vazquez Reyes C, Tangprasertchai NS, Yogesha SD, Nguyen RH, Zhang X, Rajan R, Qin PZ
- Issue date: 2017 Jun