Supplementary Materials

CRISPR-mediated live imaging of genome editing and transcription

Haifeng Wang, Muneaki Nakamura, Timothy R. Abbott, Dehua Zhao, Kaiwen Luo, Cordelia Yu, Cindy M. Nguyen, Albert Lo, Timothy P. Daley, Marie La Russa, Yanxia Liu, Lei S. Qi

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S11
  • Tables S1, S2, S4
  • Caption for Table S3
  • Captions for Movies S1 to S5
  • References
Table S3
Raw data of percentages of new and total 53BP1-colocalized Chr3 loci over time as shown in Fig. 2E-F.

Images, Video, and Other Media

Movie S1
CRISPR LiveFISH tracks dynamics of Chr13 loci in a representative normal amniotic fluid cells. Two Chr13 loci are labeled by Atto565-crRNA (gRNA, red). Hoechst 33342 (blue) labels the cell nucleus. Images were taken with seven Z confocal planes spaced by 0.2 μm every 18.5s, processed by projection of maximum intensity of all planes. Display rate 7 frames/s. See Fig. 1F.
Movie S2
CRISPR LiveFISH tracks dynamics of Chr13 loci in a representative PS patient-derived amniotic fluid cell. Three Chr13 loci are labeled by Atto565-crRNA (gRNA, red). Hoechst 33342 (blue) labels the cell nucleus. Images were taken with seven Z confocal planes spaced by 0.4 μm every 17.1s. Display rate 7 frames/s. See Fig. 1G.
Movie S3
CRISPR LiveFISH enables multi-loci chromosome imaging in living primary human T lymphocytes. Simultaneous labeling of Chr13 (Atto488-crRNA, green) and Chr3 (Cy3-crRNA, red) by co-delivery of two CRISPR RNP complexes in living primary human T cells. The recorded T lymphocyte moved slowly to the bottom right, with its movement being slowed down on a collagen pre-treated plate. Images were taken with 51 Z-confocal planes every 3 min (with a delay between frame 1&2), processed by projection of maximum intensity of all planes. Display rate 3 frames/s. See fig. S5B for another example with nucleus staining.
Movie S4
Tracking dynamics of translocated endogenous chromosomes in living U2OS cells. Simultaneous labeling of Chr3 (Atto488-gRNA, green) and Chr13 (Atto565- gRNA, red) by co-delivery of labeling gRNAs and cutting gRNAs in complex with Cas9 in living U2OS cells. A translocated chromosome is marked by a pair of closely located Chr3 and Chr13 sites in the center of nucleus. Images were taken with seven Z confocal planes spaced by 0.3 μm every 24 second, processed by projection of maximum intensity. Display rate 7 frames/s. See Fig. 3B.
Movie S5
Tracking formation of translocation between endogenous chromosomes in living U2OS cells. Simultaneous labeling of Chr3 (Atto488-gRNA, green) and Chr13 (Atto565-gRNA, red) by co-delivery of labeling gRNAs and cutting gRNAs in complex with Cas9 in living U2OS cells. Images were taken with seven Z confocal planes spaced by 0.3 μm every 0.5 hours. Display rate 4 frames/s. See Fig. 3D.