Table 2. Analysis of transposon targeting into HPRT.
| gRNA | Number of unique sites | Total sequencing reads | Reads in HPRT | % targeted into HPRT | Average # of copies/RnaseP | |
|---|---|---|---|---|---|---|
| HA-PB | N/A | 16683 | 7840617 | 1168 | 0.01 | 8.5±2.5 |
| Z3-PB | N/A | 19224 | 4959877 | 20624 | 0.42 | 36±12 |
| TAL2-PB | N/A | 21879 | 4010639 | 39054 | 0.97 | 49±10 |
| Cas9-PB | E2 | 51570 | 10288878 | 0 | 0 | ND |
| E3 | 59658 | 8956385 | 0 | 0 | ND | |
| PB + Cas9 | E2 | 25601 | 9036304 | 0 | 0 | ND |
| E3 | 64514 | 9007196 | 0 | 0 | ND |
HiSeq was used to recover thousands of unique sites using the various transposases from 6-TG resistant cells. The ‘% targeted into HPRT’ represents the sequencing reads of confirmed integrations with the cognate TTAA target site recovered between the 5′ and 3′ untranslated regions of the full genomic sequence of human HPRT in BLAT. The percentage represents the depth of coverage of recovered integration sites within HPRT divided by the total number of integrations recovered. The ‘Average # of copies/RnaseP’ represents the number of copies of the neomycin resistance gene (contained within the transposon) divided by the copies of RnaseP (number of copies of transposon per haploid genome) measured using qPCR.