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Editorial
. 2016 Dec 6;7(49):80103-80104.
doi: 10.18632/oncotarget.13371.

CRISPR-mediated multiplexed genetic manipulation

Lin Lin, Johan Vad-Nielsen, Yonglun Luo
Editorial

CRISPR-mediated multiplexed genetic manipulation

Lin Lin et al. Oncotarget. .
No abstract available

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Figures

Figure 1
Figure 1. Schematic illustration of simultaneous and orthogonal gene inhibition, activation, and knockout using Cas9 orthologues and multiplexed gRNA expression array
Six modular gRNA expression plasmids, with two gRNAs for each Cas9 orthologs (SpCas9, SaCas9, and NmCas9), are assembled into one multiplexed gRNA expression array based on Bsal-mediated Golden- Gate Assembly. Letter “B” and arrows indicated the Bsal recognition site and cleavage position, respectively. Co-delivery of the multiplexed gRNA expression array with the nuclease-deficient SpCas9 fused to transcriptional activation domain VP64 (dSpCas9-VP64), nuclease-deficient SaCas9 fused to transcriptional repression domain KRAB (dSaCas9-KRAB), and wild type NmCas9 enables simultaneous and orthogonal activation of gene A., inhibition of gene B., and knockout of gene C. Asterisks (*) represent nuclease-deficient mutations.
See this image and copyright information in PMC

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References

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