Part collection

This year, we designed, built, and characterized a set of basic parts related to transcriptional repression, fluorescent tagging, and competitive inhibition. These parts were assembled to study various proteins such as DUX4-DBD and the KRAB repressor. All parts have been submitted to The iGEM Parts Registry, following the BioBricks assembly standard. On this page, we provide a brief description of our parts, divided into several sections according to the modules they worked in within our project. For more detailed information about our parts, we recommend visiting the corresponding parts page on The iGEM Parts Registry or our Design and Engineering wiki pages.

Basic Parts

Part Collection (Basic)
Module Type Designer Code Name Length (bp) Function and Description
Gene Regulation Repressor Research Team BBa_K5195001 KRAB Repressor 195 Served as another inhibitor of DUX4-FL binding to reduce downstream effects. The KRAB domain was sourced from pHAGE EF1α dCas9-KRAB plasmid, synthetically constructed, and used in HEK 293T cells to reduce DUX4-FL expression. [1] Reference.
Coding BBa_K5195002 DUX4-DBD 651 A short isoform of DUX4-FL designed to outcompete full-length DUX4 at binding sites, reducing its downstream effects in muscular dystrophy research. [2] Reference.
Reporter BBa_K5195004 mNeonGreen 708 Green fluorescent signal utilized for tagging DBD and DBD-KRAB in cells and imaging via microscopy. mNeonGreen enhances signal intensity compared to GFP, with 506 nm excitation and 517 nm emission. It is fused with DUX4-DBD using a P2A linker for easy tracking in experiments. [2] Reference.

Composite Parts

Part Collection (Composite)
Module Type Designer Code Name Length (bp) Function and Description
Gene Regulation Measurement Research Team BBa_K5195010 imNG-P2A-DUX4-DBD 1425 Composite coding sequence containing mNeonGreen fused to DUX4-DBD with a P2A linker for quantifying DUX4-DBD via fluorescent microscopy and plate reader assay
Gene Regulation Measurement Research Team BBa_K5195012 imNG-P2A-DUX4-DBD-KRAB 1620 Composite coding sequence containing mNeonGreen fused to DUX4-DBD with a P2A linker for quantifying DUX4-DBD via fluorescent microscopy and plate reader assay