CRISPR-Cas9 off-targeting assessment with nucleic acid duplex energy parameters

CRISPR-Cas9 off-targeting assessment with nucleic acid duplex energy parameters

Collected and generated data for the paper

## Data Tables

Off-target score data for the ROC analysis using Haeussler dataset [2].

Data from the table below is used to generate the Figure-2, Table-1 and Supplementary Figure-1 in the corresponding paper [1]. Don't forget to cite the corresponding studies as well if you use this table.

• Haeussler_mm6_scores.csv.gz: This table includes the off-targeting scores of 1167036 off-target sequences, computed with CRISPRoff[1], CCTop[3], CFD[4], Cropit[5], Elevation (Elevation-score)[6], MIT[2,7] and VfoldCAS[8] methods. Off-target data has been taken from the Haeussler dataset [2].

Analysis with CIRCLE-seq dataset [9]

Data in all the three tables below has been generated to analyze the CIRCLE-seq dataset [9]. This data is further used to generate the Figure-3, Figure-4, and Supplementary Figure-4 in the corresponding paper. Don't forget to cite the corresponding studies as well if you use these tables.

• CIRCLEseq_known_off_scores.csv.gz: This table is used when generating the Figure-3 in the paper. It includes the 7 different off-targeting scores of CIRCLE-seq reported off-target sequences and the read counts from CIRCLE-seq experiments.

• CIRCLEseq_mm6_off_scores.csv.gz: This table is used when generating the Figure-4 in the paper. It includes the 7 different off-targeting scores of RIsearch2(v2.1)[10] based off-target predictions for CIRCLE-seq gRNAs.

• CIRCLEseq_specificities.csv.gz: This table is used when generating the Supplementary Figure-4 in the supplementary document of the paper. It includes the specificty scores of CIRCLE-seq gRNAs, computed with CRISPRspec[1], MIT[2,7], MIT*[1,2,7] and Elevation (Elevation-aggregate)[6] methods.

Analysis with SITE-seq dataset [11]

Data in all the three tables below has been generated to analyze the SITE-seq dataset [11]. This data is further used to generate the Figure-5, Supplementary Figure-2 and Supplementary Figure-3 in the corresponding paper. Don't forget to cite the corresponding studies as well if you use these tables.

• SITEseq_known_off_scores.csv.gz: This table is used when generating the Supplementary Figure-2 in the supplementary document of the paper. It includes the 7 different off-targeting scores of SITE-seq reported off-target sequences and the read counts from SITE-seq experiments.

• SITEseq_mm6_off_scores.csv.gz: This table is used when generating the Supplementary Figure-3 in the supplementary document of the paper. It includes the 7 different off-targeting scores of RIsearch2(v2.1) based off-target predictions for SITE-seq gRNAs.

• SITEseq_specificities.csv.gz: This table is used when generating the Figure-5 in the paper. It includes the 4 different specificty scores of SITE-seq gRNAs.

Specificity-Efficiency Analysis

This data is used to generate the Figure-6 and Supplementary Figure-5 in the corresponding paper. Don't forget to cite the corresponding studies as well if you use these tables.

• Doench_Wang_specificity_grps.csv.gz: This table includes the specificity group of 3802 gRNA/on-target sequences, computed with CRISPRspec and MIT methods. gRNA sequence and modulation frequency data have been taken from the Haeussler dataset [2].

## Citation

If you find this data useful for your research, please cite the following works where appropriate:

1. [Our citation comes here]
2. Haeussler, M., Schonig, K., Eckert, H., Eschstruth, A., Mianne, J., Renaud, J.B., Schneider-Maunoury, S., Shkumatava, A., Teboul, L., Kent, J., Joly, J.S., Concordet, J.P.: Evaluation of off-target and on-target scoring algorithms and integration into the guide RNA selection tool CRISPOR. Genome Biol. 17(1), 148 (2016). PMID 27380939
3. Stemmer, M., Thumberger, T., Del Sol Keyer, M., Wittbrodt, J., Mateo, J.L.: CCTop: An Intuitive, Flexible and Reliable CRISPR/Cas9 Target Prediction Tool. PLoS ONE 10(4), 0124633 (2015). PMID 25909470
4. Doench, J.G., Fusi, N., Sullender, M., Hegde, M., Vaimberg, E.W., Donovan, K.F., Smith, I., Tothova, Z., Wilen, C., Orchard, R., Virgin, H.W., Listgarten, J., Root, D.E.: Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nat. Biotechnol. 34(2), 184–191 (2016). PMID 26780180
5. Singh, R., Kuscu, C., Quinlan, A., Qi, Y., Adli, M.: Cas9-chromatin binding information enables more accurate CRISPR off-target prediction. Nucleic Acids Res. 43(18), 118 (2015). PMID 26032770
6. Listgarten, J., Weinstein, M., Kleinstiver, B.P., Sousa, A.A., Joung, J.K., Crawford, J., Gao, K., Hoang, L., Elibol, M., Doench, J.G., Fusi, N.: Prediction of off-target activities for the end-to-end design of CRISPR guide RNAs. Nature Biomedical Engineering 2, 38–47 (2018). PMID 29998038
7. Hsu, P.D., Scott, D.A., Weinstein, J.A., Ran, F.A., Konermann, S., Agarwala, V., Li, Y., Fine, E.J., Wu, X., Shalem, O., Cradick, T.J., Marraffini, L.A., Bao, G., Zhang, F.: DNA targeting specificity of RNA-guided Cas9 nucleases. Nat. Biotechnol. 31(9), 827–832 (2013). PMID 23873081
8. Xu, X., Duan, D., Chen, S.J.: CRISPR-Cas9 cleavage efficiency correlates strongly with target-sgRNA folding stability: from physical mechanism to off-target assessment. Sci Rep 7(1), 143 (2017). PMID 28273945
9. Tsai, S.Q., Nguyen, N.T., Malagon-Lopez, J., Topkar, V.V., Aryee, M.J., Joung, J.K.: CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR-Cas9 nuclease off-targets. Nat. Methods 14(6), 607–614 (2017). PMID 28459458
10. Alkan, F., Wenzel, A., Palasca, O., Kerpedjiev, P., Rudebeck, A.F., Stadler, P.F., Hofacker, I.L., Gorodkin, J.: RIsearch2: suffix array-based large-scale prediction of RNA-RNA interactions and siRNA off-targets. Nucleic Acids Res. (2017). PMID 28108657
11. Cameron, P., Fuller, C.K., Donohoue, P.D., Jones, B.N., Thompson, M.S., Carter, M.M., Gradia, S., Vidal, B., Garner, E., Slorach, E.M., Lau, E., Banh, L.M., Lied, A.M., Edwards, L.S., Settle, A.H., Capurso, D., Llaca, V., Deschamps, S., Cigan, M., Young, J.K., May, A.P.: Mapping the genomic landscape of CRISPR-Cas9 cleavage. Nat. Methods 14(6), 600–606 (2017). PMID 28459459

## Contact

ferro@rth.dk gorodkin@rth.dk