Rapid production of SaCas9 in plant‐based cell‐free lysate for activity testing
Abstract: Cas9 nucleases have become the most versatile tool for genome editing projects in a broad range of organisms. The recombinant production of Cas9 nuclease is desirable for in vitro activity assays or the preparation of ribonucleoproteins (RNPs) for DNA‐free genome editing approaches. For the rapid production of Cas9, we explored the use of a recently established cell‐free lysate from tobacco (Nicotiana tabacum L.) BY‐2 cells. Using this system, the 130‐kDa Cas9 nuclease from Staphylococcus aureus (SaCas9) was produced and subsequently purified via affinity chromatography. The purified apoenzyme was supplemented with 10 different sgRNAs, and the nuclease activity was confirmed by the linearization of plasmid DNA containing cloned DNA target sequences.
- Location
-
Deutsche Nationalbibliothek Frankfurt am Main
- Extent
-
Online-Ressource
- Language
-
Englisch
- Bibliographic citation
-
Rapid production of SaCas9 in plant‐based cell‐free lysate for activity testing ; day:27 ; month:04 ; year:2022 ; extent:7
Biotechnology journal ; (27.04.2022) (gesamt 7)
- Creator
-
Schiermeyer, Andreas
Cerda‐Bennasser, Pedro
Schmelter, Thomas
Huang, Xin
Christou, Paul
Schillberg, Stefan
- DOI
-
10.1002/biot.202100564
- URN
-
urn:nbn:de:101:1-2022042815101780344714
- Rights
-
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Last update
-
15.08.2025, 7:29 AM CEST
Data provider
This object is provided by:
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.
Associated
- Schiermeyer, Andreas
- Cerda‐Bennasser, Pedro
- Schmelter, Thomas
- Huang, Xin
- Christou, Paul
- Schillberg, Stefan
Other Objects (12)
Persistence of bank credit default swap spreads
Stem cell-based drug delivery strategy for skin regeneration and wound healing: potential clinical applications
A Scene Unmixing Deep Learning Network for Local Climate Zone Mapping and Analysis Using Very High Resolution Remote Sensing Imagery
In vivo gene editing and in situ generation of chimeric antigen receptor cells for next-generation cancer immunotherapy
Analysis of runoff variation and driving mechanism in Huangfuchuan River Basin in the middle reaches of the Yellow River, China
Structural Engineering for High‐Performance Flexible and Stretchable Strain Sensors
Tobacco BY-2 cell-free lysate: an alternative and highly-productive plant-based in vitro translation system
Tobacco BY-2 cell-free lysate : an alternative and highly-productive plant-based in vitro translation system
The role of BCL11B in hematological malignancy
Age-related hypertrophy of adenoid and tonsil with its relationship with craniofacial morphology
Association between GDF5 rs143383 genetic polymorphism and musculoskeletal degenerative diseases susceptibility: a meta-analysis
Comparative performance of transcriptome assembly methods for non-model organisms
Persistence of bank credit default swap spreads
Stem cell-based drug delivery strategy for skin regeneration and wound healing: potential clinical applications
A Scene Unmixing Deep Learning Network for Local Climate Zone Mapping and Analysis Using Very High Resolution Remote Sensing Imagery
In vivo gene editing and in situ generation of chimeric antigen receptor cells for next-generation cancer immunotherapy
Analysis of runoff variation and driving mechanism in Huangfuchuan River Basin in the middle reaches of the Yellow River, China
Structural Engineering for High‐Performance Flexible and Stretchable Strain Sensors
Tobacco BY-2 cell-free lysate: an alternative and highly-productive plant-based in vitro translation system
Tobacco BY-2 cell-free lysate : an alternative and highly-productive plant-based in vitro translation system
The role of BCL11B in hematological malignancy
Age-related hypertrophy of adenoid and tonsil with its relationship with craniofacial morphology
Association between GDF5 rs143383 genetic polymorphism and musculoskeletal degenerative diseases susceptibility: a meta-analysis
Comparative performance of transcriptome assembly methods for non-model organisms
Persistence of bank credit default swap spreads
Stem cell-based drug delivery strategy for skin regeneration and wound healing: potential clinical applications
A Scene Unmixing Deep Learning Network for Local Climate Zone Mapping and Analysis Using Very High Resolution Remote Sensing Imagery
In vivo gene editing and in situ generation of chimeric antigen receptor cells for next-generation cancer immunotherapy
Analysis of runoff variation and driving mechanism in Huangfuchuan River Basin in the middle reaches of the Yellow River, China
Structural Engineering for High‐Performance Flexible and Stretchable Strain Sensors
Tobacco BY-2 cell-free lysate: an alternative and highly-productive plant-based in vitro translation system
Tobacco BY-2 cell-free lysate : an alternative and highly-productive plant-based in vitro translation system
The role of BCL11B in hematological malignancy
Age-related hypertrophy of adenoid and tonsil with its relationship with craniofacial morphology
Association between GDF5 rs143383 genetic polymorphism and musculoskeletal degenerative diseases susceptibility: a meta-analysis