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C-NHEJ without indels is robust and requires synergistic function of distinct XLF domains.
Bhargava R, Sandhu M, Muk S, Lee G, Vaidehi N, Stark JM
Nat Commun. 2018 Jun 27;9(1):2484. doi: 10.1038/s41467-018-04867-5.
PubMed

Plasmids from Article

ID Plasmid Purpose  
113617pCMV6-AC EJ7-GFPReporter for canonical-NHEJ. Reporter for end joining between two double-strand breaks, induced with the 7a and 7b sgRNAs with CAS9. EJ between the distal ends w/o indel mutations restores GFP
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113619pCMV6-AC 4-μHOMReporter for Alt-EJ, variant of EJ7-GFP. Double strand breaks are induced with the 7a & 7h, or 7a & 7i sgRNAs, with CAS9. EJ using 4 nt of microhomology causes a deletion mutation and restores GFP
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1136207a sgRNA for EJ7-GFP and 4-μHOM reporterssgRNA/CAS9 expression plasmid to induce the 5’ double-strand break in both the EJ7-GFP and 4-μHOM reporters
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1136247b sgRNA for EJ7-GFP reportersgRNA/CAS9 expression plasmid to induce the 3’ double-strand break in the EJ7-GFP reporter
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1136257h sgRNA for 4-μHOM reportersgRNA/CAS9 expression plasmid to induce a 3’ double-strand break in the 4-μHOM reporter at the edge of the microhomology.
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1136267i sgRNA for 4-μHOM reportersgRNA/CAS9 expression plasmid to induce a 3’ double-strand break in the 4-μHOM reporter 8 nts. upstream from the microhomology
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