Fig. 6: Tag-seq profiles the integration loci of exogenous genes induced by transposons. | Communications Biology

Fig. 6: Tag-seq profiles the integration loci of exogenous genes induced by transposons.

From: Tag-seq: a convenient and scalable method for genome-wide specificity assessment of CRISPR/Cas nucleases

Fig. 6

a Diagram showing the use of Tag-seq to discover integration sites of EGFP induced by Cas9/dCas9 and Sleeping Beauty transposase fusion proteins. Arm-L/Arm-R, the left/right homology arm sequences. UMI, unique molecular index, S.B., sample barcode. b Workflow for construction of the library of Tag-seq for detection of the integration locations induced by Sleeping Beauty transposons. c Venn diagram showing the overlapping numbers of the insertion sites among the control, Cas9-SB, and dCas9-SB by Tag-seq. Ctrl, genomic DNA from blank-transfected cells was analyzed with Tag-seq; Cas9-SB/dCas9-SB, genomic DNA from cells transfected with the plasmids coding Cas9/dCas9-SB and the donor plasmid containing the SB-transposon-arm-flanked EGFP-expression cassette was analyzed with Tag-seq. d Sequence logos obtained via DNA sequences near the cleavage sites (±5 bp) identified by Tag-seq. e Distributions of the integrated locations in genome.

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