Table 1.
Results of human sgRNA testing
| Gene | sgRNA ID | Target sequence | Experiment 1–replicate 1 | Experiment 2–replicate 1 | Experiment 2–replicate 2 | Experiment 3–replicate 1 | Experiment 3–replicate 2 | Experiment 4–replicate 1 | Experiment 4–replicate 2 | Experiment 5–replicate 1 | Experiment 5–replicate 2 | Average vs. GAPDH | SEM |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| GATA4 | sgGv1-A | GCCCAGCGGAGG TGTAGCCG |
261.083 | 69.807 | 43.950 | – | – | 234.804 | 111.798 | 701.449 | 544.367 | 281.037 | 94.880 |
| GATA4 | sgGv1-B | TAGCACTTGGGC ATTTTCCG |
21.432 | – | – | – | – | – | – | – | – | 21.432 | NA |
| GATA4 | sgGv1-C | CCTGTGGGAGTC ACGTGCAA |
5.110 | – | – | – | – | – | – | – | – | 5.110 | NA |
| GATA4 | sgGv3-1 | AGGGACCATGTA TCAGAGCTTGG |
– | – | – | – | – | – | – | 314.788 | 174.231 | 244.510 | 70.279 |
| GATA4 | sgGv3-2 | TTGACCTGCGAG GGAGAGAGAGG |
– | – | – | – | – | – | – | 10.909 | 12.211 | 11.560 | 0.651 |
| GATA4 | sgGv3-4 | GCGTGGCTCCTT GACCTGCGAGG |
– | – | – | – | – | – | – | 21.198 | 16.129 | 18.664 | 2.534 |
| GATA4 | sgGv3-5 | TCGCAGGTCAAG GAGCCACGCGG |
– | – | – | – | – | – | – | 24.016 | 107.836 | 65.926 | 41.910 |
| GATA4 | sgGv3-6 | CGCAGGTCAAGG AGCCACGCGGG |
– | – | – | – | – | – | – | 173.596 | 202.758 | 188.177 | 14.581 |
| GATA4 | sgGv4-1 | TGAGGACTGAGT GCCGCGCAGGG |
– | – | – | – | – | – | – | 34.227 | 23.207 | 28.717 | 5.510 |
| GATA4 | sgGv4-2 | GGCGGGCGGCG CAAACTCGGCGG |
– | – | – | – | – | – | – | 1.433 | 0.778 | 1.105 | 0.328 |
| GATA4 | sgGv4-3 | CAGTACTTAAGG CACATCTAAGG |
– | – | – | – | – | – | – | 0.484 | 0.527 | 0.505 | 0.021 |
| GATA4 | sgGv4-4 | AGAGAGGCAGAT GTATAGACAGG |
– | – | – | – | – | – | – | 0.747 | 0.930 | 0.839 | 0.092 |
| GATA4 | sgGv4-5 | ATAAACGGGCCA AAGGTACCTGG |
– | – | – | – | – | – | – | 0.664 | 0.522 | 0.593 | 0.071 |
| GATA4 | sgGv4-6 | ATCTTCAGTCATC AAGGATGGGG |
– | – | – | – | – | – | – | 0.793 | 0.757 | 0.775 | 0.018 |
| GATA4 | sgGv4-7 | ATGTGATGCCCAG AGTCAGTGGG |
– | – | – | – | – | – | – | 0.581 | 0.836 | 0.708 | 0.128 |
| GATA4 | sgGv4-8 | CTTCCTACGATGT TTATGACTGG |
– | – | – | – | – | – | – | 0.495 | 0.562 | 0.528 | 0.034 |
| GATA4 | sgGv4-9 | AATTAAATACATC TACTGAGAGG |
– | – | – | – | – | – | – | 0.612 | 0.495 | 0.553 | 0.058 |
| GATA4 | sgGv4-10 | TCAGCATTTATCC GGAGCGGGGG |
– | – | – | – | – | – | – | 0.758 | 0.911 | 0.835 | 0.077 |
| MEF2C | sgMv1-A | TATCAAGGAAAT AAACTATA |
1.133 | – | – | – | – | – | – | – | – | 1.133 | NA |
| MEF2C | sgMv1-B | ATATATGGTATA TCACAGAC |
0.873 | – | – | – | – | – | – | – | – | 0.873 | NA |
| MEF2C | sgMv1-R1A | TCAAGGAGGTAC AAAAGGAA |
0.872 | – | – | – | – | – | – | – | – | 0.872 | NA |
| MEF2C | sgMv1-R3A | GATGAAATGAAA AACTGGAG |
1.592 | – | – | – | – | – | – | – | – | 1.592 | NA |
| MEF2C | sgMv1-R2A | TCGTGCCAAGGA AAATGGAG |
0.912 | – | – | – | – | – | – | – | – | 0.912 | NA |
| MEF2C | sgMv1-R2B | AAAGTACAGTGG GCTAACAG |
0.694 | – | – | – | – | – | – | – | – | 0.694 | NA |
| MEF2C | sgMv2-9 | CCTCCTCCTCCTC TTCCGAG |
– | – | – | 11.601 | 16.600 | – | – | – | – | 14.101 | 2.500 |
| MEF2C | sgMv2-10 | CCGAGGCCGCTC GGAAGAGG |
– | – | – | 57.353 | 62.174 | 96.684 | 98.050 | 44.579 | 53.283 | 68.687 | 9.373 |
| MEF2C | sgMv2-15 | CCTCGGCGCGCG CGAATGCG |
– | – | – | 0.734 | 0.959 | – | – | – | – | 0.847 | 0.112 |
| MEF2C | sgMv2-17 | CCGCGCATTCGC GCGCGCCG |
– | – | – | 0.672 | 0.708 | – | – | – | – | 0.690 | 0.018 |
| MEF2C | sgMv3-1 | AGCTGTGCAAGTG CTGAAGAAGG |
– | – | – | – | – | 43.275 | 64.776 | – | – | 54.026 | 10.750 |
| MEF2C | sgMv3-3 | ATATATGGTATATC ACAGACAGG |
– | – | – | – | – | 2.427 | 2.701 | – | – | 2.564 | 0.137 |
| MEF2C | sgMv3-4 | GTACCTGTGTATG CTAAGTAGGG |
– | – | – | – | – | 2.849 | 2.696 | – | – | 2.772 | 0.077 |
| MEF2C | sgMv3-6 | GCAGCTAATTCA TTTCTGAAGGG |
– | – | – | – | – | 1.633 | 1.735 | – | – | 1.684 | 0.051 |
| MEF2C | sgMv3-8 | CCATCCTATTTG CATAACGAGGG |
– | – | – | – | – | 1.155 | 1.421 | – | – | 1.288 | 0.133 |
| MEF2C | sgMv3-9 | CCCTCGTTATGC AAATAGGATGG |
– | – | – | – | – | 1.862 | 2.055 | – | – | 1.958 | 0.097 |
| MEF2C | sgMv3-10 | TCCATCCTATTT GCATAACGAGG |
– | – | – | – | – | 1.735 | 2.119 | – | – | 1.927 | 0.192 |
| MEF2C | sgMv3-11 | ACAGACATATCA TGTCTTTGTGG |
– | – | – | – | – | 2.609 | 3.066 | – | – | 2.837 | 0.229 |
| MEF2C | sgMv3-12 | ATTACATACTGG AGATCTCTGGG |
– | – | – | – | – | 2.058 | 2.641 | – | – | 2.350 | 0.291 |
| MEF2C | sgMv3-13 | ATACTGGAGATC TCTGGGTCAGG |
– | – | – | – | – | 4.082 | 3.801 | – | – | 3.941 | 0.140 |
| MEF2C | sgMv3-14 | GAAGAAAAACGG GGACTATGGGG |
– | – | – | – | – | 1.167 | 1.719 | – | – | 1.443 | 0.276 |
| MEF2C | sgMv4-1 | CTGGTCTGGCTT TATTCTGCAGG |
– | – | – | – | – | – | – | 0.859 | 0.786 | 0.823 | 0.037 |
| MEF2C | sgMv4-2 | TGCCATTTTACA TGTTCCAGTGG |
– | – | – | – | – | – | – | 1.075 | 0.841 | 0.958 | 0.117 |
| MEF2C | sgMv4-3 | CACTTATTCAGTA CTCAGCTAGG |
– | – | – | – | – | – | – | 1.424 | 1.281 | 1.353 | 0.071 |
| MEF2C | sgMv4-4 | GGTTTATTGGTAA ACATGTATGG |
– | – | – | – | – | – | – | 1.155 | 1.144 | 1.150 | 0.005 |
| MEF2C | sgMv4-5 | ATATTACTTTTCA TCGAGGGTGG |
– | – | – | – | – | – | – | 0.904 | 0.852 | 0.878 | 0.026 |
| MEF2C | sgMv4-6 | CTACCAAACTCCA CTTTCAGTGG |
– | – | – | – | – | – | – | 1.116 | 1.354 | 1.235 | 0.119 |
| MEF2C | sgMv4-7 | CCAGAGCTAGGT TGTTTATGTGG |
– | – | – | – | – | – | – | 1.067 | 1.229 | 1.148 | 0.081 |
| MEF2C | sgMv4-8 | ATGGTGTGGTCT CACCTTACTGG |
– | – | – | – | – | – | – | 0.940 | 0.962 | 0.951 | 0.011 |
| MEF2C | sgMv4-9 | CTTGTCCAAGGT TCTGCCAAAGG |
– | – | – | – | – | – | – | 1.140 | 1.408 | 1.274 | 0.134 |
| MEF2C | sgMv4-10 | GACAAATACATA CTAATTTGAGG |
– | – | – | – | – | – | – | 1.041 | 1.036 | 1.038 | 0.003 |
| TBX5 | sgTv1-A | CAAATCATTTCA AACGCGGT |
13.794 | – | – | – | – | – | – | – | – | 13.794 | NA |
| TBX5 | sgTv1-B | CTGCTTCTCTGA GCAAGCGG |
20.402 | – | – | – | – | – | – | – | – | 20.402 | NA |
| TBX5 | sgTv1-C | TGAGTGTTCGG GAAATCTAG |
10.990 | – | – | – | – | – | – | – | – | 10.990 | NA |
| TBX5 | sgTv2-12 | GACGTCACGAG TCACGCAAC |
– | – | – | 0.122 | 0.086 | – | – | – | – | 0.104 | 0.018 |
| TBX5 | sgTv2-15 | GCGCTGTCACG TTTGGCTGG |
– | 0.511 | 2.218 | 0.309 | 0.485 | – | – | – | – | 0.881 | 0.448 |
| TBX5 | sgTv2-34 | GCCGCTGCACC TATAGGACT |
– | – | – | 0.141 | 0.113 | – | – | – | – | 0.127 | 0.014 |
| TBX5 | sgTv2-35 | AAACGTGACAG CGCGCGGGC |
– | – | – | 0.751 | 0.594 | – | – | – | – | 0.673 | 0.079 |
| TBX5 | sgTv3-1 | CACCATGGCCG ACGCAGACGAGG |
– | – | – | – | – | 6,851.464 | 4,704.819 | – | – | 5,778.141 | 1,073.323 |
| TBX5 | sgTv3-2 | CTCAGAGCAGA ACCTTGCGCGGG |
– | – | – | – | – | 10,238.454 | 9,834.440 | – | – | 10,036.447 | 202.007 |
| TBX5 | sgTv3-4 | CCTCAGAGCAG AACCTTGCGCGG |
– | – | – | – | – | 13,035.915 | 12,940.372 | 16.275 | 15.789 | 6,502.088 | 3,744.777 |
| TBX5 | sgTv3-5 | TGCTCTGAGGAC AAGAAGCAGGG |
– | – | – | – | – | 1,843.998 | 1,637.516 | – | – | 1,740.757 | 103.241 |
| TBX5 | sgTv3-8 | GCACAATTCTAGT GACAGGAGGG |
– | – | – | – | – | 30,360.732 | 512.313 | – | – | 15,436.522 | 14,924.209 |
| TBX5 | sgTv3-9 | AGTGACAGGAGGGA GCAGTTTGG |
– | – | – | – | – | 381.584 | 418.221 | – | – | 399.903 | 18.318 |
| TBX5 | sgTv3-10 | TGCTGGTAGCTGGA AACTGGGGG |
– | – | – | – | – | 6.473 | 22.619 | – | – | 14.546 | 8.073 |
| TBX5 | sgTv3-11 | GCACCTCCGTTCCC AGCCGAAGG |
– | – | – | – | – | 3,237.790 | 2,387.540 | – | – | 2,812.665 | 425.125 |
| TBX5 | sgTv3-12 | TAGTAACAGTAAT AATTCTAGGG |
– | – | – | – | – | 98.805 | 80.128 | – | – | 89.467 | 9.338 |
| TBX5 | sgTv3-13 | AAAGTAATAGTAA TACTACTAGG |
– | – | – | – | – | 11.146 | 14.097 | – | – | 12.622 | 1.475 |
| TBX5 | sgTv3-14 | ACCCGCGTCAGAC CCGGAGAAGG |
– | – | – | – | – | 49.323 | 50.656 | – | – | 49.990 | 0.666 |
| TBX5 | sgTv3-16 | ATCACGGAGAGCC TGCGCAGGGG |
– | – | – | – | – | 6,474.837 | 6,722.309 | – | – | 6,598.573 | 123.736 |
| TBX5 | sgTv3-17 | TATCACGGAGAGC CTGCGCAGGG |
– | – | – | – | – | 1,998.376 | 2,470.274 | – | – | 2,234.325 | 235.949 |
| TBX5 | sgTv3-18 | TTATCACGGAGAG CCTGCGCAGG |
– | – | – | – | – | 651.057 | 946.434 | – | – | 798.745 | 147.688 |
| TBX5 | sgTv3-19 | GACGGGCAGCTTG CTTATCACGG |
– | – | – | – | – | 2,752.702 | 3,050.598 | – | – | 2,901.650 | 148.948 |
| TBX5 | sgTv3-20 | GAAGTCCAGACAA ATTTGACGGG |
– | – | – | – | – | 121.863 | 170.663 | – | – | 146.263 | 24.400 |
| TBX5 | sgTv3-21 | AGAAGTCCAGACA AATTTGACGG |
– | – | – | – | – | 1.558 | 3.550 | – | – | 2.554 | 0.996 |
| TBX5 | sgTv3-26 | ATTTTTGGAAGCC ACTGGGTAGG |
– | – | – | – | – | 691.631 | 780.797 | – | – | 736.214 | 44.583 |
| TBX5 | sgTv3-28 | GAAATTCTCTAA AGGACACTGGG |
– | – | – | – | – | 14.393 | 13.128 | – | – | 13.761 | 0.633 |
| TBX5 | sgTv3-29 | AGAAATTCTCTAA AGGACACTGG |
– | – | – | – | – | 4.952 | 3.883 | – | – | 4.417 | 0.535 |
| TBX5 | sgTv3-30 | AGAAAACTATTCC TCTTGTGAGG |
– | – | – | – | – | 44.358 | 56.180 | – | – | 50.269 | 5.911 |
| TBX5 | sgTv4-1 | GAGGTCTCTTGCA TAAGGCATGG |
– | – | – | – | – | – | – | 0.448 | 0.465 | 0.456 | 0.009 |
| TBX5 | sgTv4-2 | AACCAGCTAGAGC GGCGCCTCGG |
– | – | – | – | – | – | – | 0.253 | 0.628 | 0.441 | 0.188 |
| TBX5 | sgTv4-3 | CTTGGACCTTTCT CTCCGCAAGG |
– | – | – | – | – | – | – | 1.031 | 0.776 | 0.903 | 0.127 |
| TBX5 | sgTv4-4 | AAATAGAGTGCCT CGTGCCTCGG |
– | – | – | – | – | – | – | 0.290 | 0.514 | 0.402 | 0.112 |
| TBX5 | sgTv4-5 | GTTTTGCAACTCA ACCTAGTGGG |
– | – | – | – | – | – | – | 0.388 | 0.411 | 0.400 | 0.012 |
| TBX5 | sgTv4-6 | AGAATCGAACCCC GAAGCTGGGG |
– | – | – | – | – | – | – | 0.564 | 0.234 | 0.399 | 0.165 |
| TBX5 | sgTv4-7 | CATGCTTCGGAAA GCCCTGGGGG |
– | – | – | – | – | – | – | 2.066 | 1.102 | 1.584 | 0.482 |
| TBX5 | sgTv4-8 | GAAGCTGCCCGCT CTCCCCGCGG |
– | – | – | – | – | – | – | 9.083 | 7.181 | 8.132 | 0.951 |
| TBX5 | sgTv4-9 | TGGGAAACGAAAG CGAAGCTCGG |
– | – | – | – | – | – | – | 7.626 | 9.684 | 8.655 | 1.029 |
| TBX5 | sgTv4-10 | AAACACGAGCCTC GGATTTGGGG |
– | – | – | – | – | – | – | 8.480 | 10.090 | 9.285 | 0.805 |
GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NA, not applicable.