Thermostable Cas9 nucleases with reduced off-target activity

ThermoCas9 is identified and characterized from the thermophilic bacterium Geobacillus thermodenitrificans T12. Experiments show how in vitro ThermoCas9 is active between 20 and 70 °C, has stringent PAM-preference at lower temperatures, tolerates fewer spacer-protospacer mismatches than SpCas9 and its activity at elevated temperatures depends on the sgRNA-structure. Described are ThermoCas9-based engineering tools for gene deletion and transcriptional silencing at 5 °C in Bacillus smithii and for gene deletion at 37 °C in Pseudomonas putida

Bibliographic Details

Main Authors:	van der Oost, J., van Kranenburg, R., Bosma, E.F., Mougiakos, I., Mohanraju, P.
Format:	Patent biblioteca
Language:	English
Subjects:	Life Science,
Online Access:	https://research.wur.nl/en/publications/thermostable-cas9-nucleases-with-reduced-off-target-activity
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