CC-5707 fancd2 [line #1]

$30.00

From Chew Yen Peng, Aron Ferenczi and Attila Molnar, University of Edinburgh-UK, December 2021

Knock-out of fancd2 (Cre10.g453650) in CC-1883, resulting in deficiency of Fanconi anaemia (FA) DNA repair. One of three fancd2 lines deposited (CC-5707, CC-5708, CC-5709).

Knock-out generated using CRISPR/Cas12a and the 1.8 kb aphVIII antibiotic selection marker to disrupt the gene of interest, resulting in paromomycin resistance (paroR).

FA deficiency was confirmed by hypersensitivity to the DNA-damaging agent mitomycin C, and the knock-out locus was sequence-verified (see Ferenczi et al. 2021, Supplementary Fig. 9). No off-target analysis was performed; may contain off-target edits and aphVIII integrations.

For the Cas12a gRNA target sequence and PCR conditions (incl. primers), see Ferenczi et al. 2021, Supplementary Data 7 and 9, respectively. Up to 10% DMSO may be added to the PCR to help amplify long GC-rich amplicons, in which case lower annealing temperatures by 3–5°C.

Deposited as part of 21 lines containing eight different DNA repair mutant genotypes (CC-5701 to CC-5721). For full list, see Ferenczi et al. 2021, Supplementary Data 10.

Ferenczi A, Chew YP, Kroll E, von Koppenfels C, Hudson A, Molnar A. Mechanistic and genetic basis of single-strand templated repair at Cas12a-induced DNA breaks in Chlamydomonas reinhardtii. Nat Commun. 2021 Nov 19;12(1):6751. doi: 10.1038/s41467-021-27004-1. PMID: 34799578; PMCID: PMC8604939.