(i) AAV2 plasmids. Plasmids pAV2Rep, pCMVAV2Cap, pAAVRep2Cap2 (pR2C2), and pAV2F5tg83luc-CMVmCherry (4.6-kb) have been described previously (35 (link), 76 (link)). AAV2 infectious clone SSV9 (pSub201[-]), containing a full-length AAV2 genome (77 (link)), was a gift from Dr. R. J. Samulski at the University of North Carolina, Chapel Hill (78 (link)).
(ii) Ad plasmids. pAd5E1 cloned with the first 4,031 bp of the Ad5 genome (containing E1 gene) was purchased from OD260 Inc., Boise, ID (#QP-04/pAd1127). pHelper plasmid was purchased from Agilent Technologies, Inc., Santa Clara, CA (#240071).
(iii) pLKO-shRNA constructs. The shRNA-expressing constructs, pLKO-shScram, pLKO-shATM, pLKO-shATR, and pLKO-shDNA-PKcs have been previously described (31 (link)).
(iv) pLentiCRISPRv2 constructs. Three sgRNAs targeting Pol η were cloned in 3 plentiCRISPRv2-Pol η KO vectors (Pol η-1: 5′-GCA CAA GTT CGT GAG TCC CG-3′; Pol η-2: 5′-GAC TGA CCC ATG TGA AAC CA-3′; Pol η-3: 5′-CTG CTC CCA CGG TGA GCT GC-3′). The 3 sgRNAs targeting Pol κ in lentiCRISPRv2-Pol κ KO are Pol κ-1: 5′-TAG GTT CAA CAC ACC TGA CG-3′, Pol κ-2: 5′-ATC CAT GTC AAT GTG CAC TA-3′, and Pol κ-3: 5′-CTT CTC CTT TGT GCT ATC CA-3′. The lentiCRISPRv2-SsgRNA expresses a scramble sgRNA (SsgRNA), containing a random sequence of 5′-GTA TTA CTG ATA TTG GTG GG-3′.
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