# name
specificity profiles of TALENs targeting CCR5A

# characteristics.description[].text
Sequences from Guilinger, et al. Nature Methods, 2014. The specificity profiles of TALENs targeting CCR5A target sites using an in vitro selection method. Briefly, preselection libraries of > 1000000000000 DNA sequences, each theoretically containing at least 10 copies of all possible DNA sequences with six or fewer mutations relative to the on-target sequence (TTCATTACACCTGCAGCT-random spacer-AGTATCAATTCTGGAAGA), were digested with 4 nM to 32 nM of an in vitro-translated TALEN. Cleaved library members harbored a free 5' monophosphate that enabled them to be captured by adaptor ligation. We isolated DNA fragments of length corresponding to 1.5 target sites (an intact target site and a repeated half-site up to the point of TALEN-induced DNA cleavage) by gel purification. High-throughput sequencing of TALEN-treated or control samples that survived this selection revealed the abundance of all TALEN-cleaved sequences as well as the abundance of the corresponding sequences before selection. In the control sample, all members of the preselection library were cleaved by a restriction endonuclease at a constant sequence to enable their capture by adaptor ligation and isolation by gel purification. Selections were indexed with the first 5 bp of sequence. Thus, the first 5bp of each sequences identifies which selection condition the sequence originated from. The indexing is as follows (with selection names corresponding to conditions given in Supplementary Table 1): for sequences in CCR5TALset1_S9 files: ACTGT = CCR5A 32 nM canonical, CTGAA = CCR5A 16 nM canonical, TGACT = CCR5A 8 nM canonical, TGCAA = CCR5A 4 nM canonical, GCATT = CCR5A 32 nM Q7, CATGA = CCR5A 16 nM Q7, ATGCT = CCR5A 8 nM Q7, CTAGT = CCR5A 4 nM Q7, GCTAA = CCR5A Q3, CAGTT = CCR5A 28-aa, GTCAT = CCR5A canonical ELD/KKR, ACGTA = CCR5A Q3 ELD/KKR. For sequences in CCR5TALset2_S10 files: ACTGT = CCR5A N1, CTGAA = CCR5A N2, TGACT = CCR5A N3, CATGA = CCR5A N2 ELD/KKR, ACGTA = CCR5A Q7 ELD/KKR Pre-selection library sequences are in CCR5ALib_S11 files