Part:BBa_K5062025

DR2

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Overview

DR2 is the second spacer of the gRNA array, placed between the PD-1 gRNA and the VTCN1 gRNA. The design is optimized to minimize recombination occurrence (Anvar et al., 2024), and the sequence is derived from the top performers from (DeWeirdt et al., 2020)

References

Anvar, N. E., Lin, C., Ma, X., Wilson, L. L., Steger, R., Sangree, A. K., Colic, M., Wang, S. H., Doench, J. G., & Hart, T. (2024). Efficient gene knockout and genetic interaction screening using the in4mer CRISPR/Cas12a multiplex knockout platform. Nature Communications, 15(1). https://doi.org/10.1038/s41467-024-47795-3

DeWeirdt, P. C., Sanson, K. R., Sangree, A. K., Hegde, M., Hanna, R. E., Feeley, M. N., Griffith, A. L., Teng, T., Borys, S. M., Strand, C., Joung, J. K., Kleinstiver, B. P., Pan, X., Huang, A., & Doench, J. G. (2020). Optimization of AsCas12a for combinatorial genetic screens in human cells. Nature Biotechnology, 39(1), 94–104. https://doi.org/10.1038/s41587-020-0600-6