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.
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PubMed
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 113617 | pCMV6-AC EJ7-GFP | Reporter 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 |
| 113619 | pCMV6-AC 4-μHOM | Reporter 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 |
| 113620 | 7a sgRNA for EJ7-GFP and 4-μHOM reporters | sgRNA/CAS9 expression plasmid to induce the 5’ double-strand break in both the EJ7-GFP and 4-μHOM reporters |
| 113624 | 7b sgRNA for EJ7-GFP reporter | sgRNA/CAS9 expression plasmid to induce the 3’ double-strand break in the EJ7-GFP reporter |
| 113625 | 7h sgRNA for 4-μHOM reporter | sgRNA/CAS9 expression plasmid to induce a 3’ double-strand break in the 4-μHOM reporter at the edge of the microhomology. |
| 113626 | 7i sgRNA for 4-μHOM reporter | sgRNA/CAS9 expression plasmid to induce a 3’ double-strand break in the 4-μHOM reporter 8 nts. upstream from the microhomology |