Application of Cas12j for Streptomyces editing and cluster activation

Tan, L. L.; Heng, E.; Zulkarnain, N.; Leong, C. Y.; Ng, V.; Yang, L. K.; Seow, D. C. S.; Peh, G.; Lim, Y. H.; Lokanand, K.; Kanagasundaram, Y.; Ng, S. B.; Wong, F. T.

doi:10.1101/2021.10.28.465406

bioRxiv · 10.1101/2021.10.28.465406

Application of Cas12j for Streptomyces editing and cluster activation

Tan, L. L.·Heng, E.·Zulkarnain, N.·Leong, C. Y.·Ng, V.·Yang, L. K.·Seow, D. C. S.·Peh, G.·Lim, Y. H.·Lokanand, K.·Kanagasundaram, Y.·Ng, S. B.·Wong, F. T.

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Abstract

In recent years, CRISPR-Cas toolboxes for Streptomyces editing have rapidly accelerated natural product discovery and engineering. However, Cas efficiencies are also oftentimes strain dependent, subsequently a variety of Cas proteins would allow for flexibility and enable genetic manipulation within a wider range of Streptomyces strains. In this work, we have further expanded the Cas toolbox by presenting the first example of Cas12j mediated editing in Streptomyces sp. A34053. In our study, we have also observed significantly improved editing efficiencies with Acidaminococcus sp. Cas12j compared to Cas12a, Francisella tularensis subsp. novicida U112s type V-A Cas (FnCpf1).

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Tan, L. L., Heng, E., Zulkarnain, N., Leong, C. Y., Ng, V., Yang, L. K., Seow, D. C. S., Peh, G., Lim, Y. H., Lokanand, K., Kanagasundaram, Y., Ng, S. B., Wong, F. T.. 2021-10-29. Application of Cas12j for Streptomyces editing and cluster activation. https://doi.org/10.1101/2021.10.28.465406

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