Emily Cliff, PhD

Synthetic Biologist

CRISPR-Based Gene Repositioning


Figure 1. Genomic sites can be physically repositioned by fusing a DNA binding domain to a membrane protein. The DNA binding domain in this instance is a CRISPR-Cas complex that can be programmed to different target sites. The CRISPR-Cas complex can be linked to a membrane protein via a scaffold RNA (scRNA), a modified gRNA that includes an MS2 RNA hairpin to recruit the MS2 coat protein (MCP)  (see Zalatan et al., 2015 for details on scaffold RNA recruitment strategy). MCP is fused to the membrane protein, thus recruiting the gene to the nuclear membrane.
Citations
  1.   Zalatan, J. G. et al. Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell  160, 339–350 (2015).
  2.   Cliff, E. R. et al. CRISPR-Cas-Mediated Tethering Recruits the Yeast HMR Mating-Type Locus to the Nuclear Periphery but Fails to Silence Gene Expression. ACS Synth. Biol. 10, 2870–2877 (2021).