An intersectional expression platform for gene complementation using RNA-fragment end joining (REJ)
- 1Salk Institute for Biological Studies
Abstract
Gene complementation is a powerful tool for genetic selection and protein functional studies but typically requires extensive screening for complementary components. Here we engineered a platform for gene complementation based on RNA end-joining (REJ) that precisely and efficiently splices separate RNA units into functional coding mRNAs. REJ is mediated by short structured modular RNA segments of ~250bp that: (a) promote RNA::RNA interaction, (b) coopt the intrinsic cell splicing machinery to facilitate RNA trans-splicing, (c) minimizes translation of protein fragments from un-spliced RNA segments, and (d) encodes scar-free protein. We demonstrate that REJ is a broadly applicable system that can reliably split nearly any gene into complementary segments regardless of protein structure. To enable the use of this system we provide a toolbox of reporters and a web-based design tool to facilitate vector design. Demonstrated REJ applications include genetic complementation, intersectional labeling of cell types, and efficient expression of large proteins.
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BibTeX
@misc{bachmann2026intersectional,
title = {{An intersectional expression platform for gene complementation using RNA-fragment end joining (REJ)}},
author = {L. C. Bachmann and R. H. Hsu and K. J. Hermann and C. E. Williams and R. L. Farman and N. Criales and C. Clark and S. Kramer and C. Thorn Perez and S. Randles and K. Lettieri and S. L. Pfaff},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.13.751279},
url = {https://www.biorxiv.org/content/10.64898/2026.09.13.751279v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - An intersectional expression platform for gene complementation using RNA-fragment end joining (REJ)
AU - L. C. Bachmann
AU - R. H. Hsu
AU - K. J. Hermann
AU - C. E. Williams
AU - R. L. Farman
AU - N. Criales
AU - C. Clark
AU - S. Kramer
AU - C. Thorn Perez
AU - S. Randles
AU - K. Lettieri
AU - S. L. Pfaff
PY - 2026
DA - 2026/09/13
PB - bioRxiv
DO - 10.64898/2026.09.13.751279
UR - https://www.biorxiv.org/content/10.64898/2026.09.13.751279v1
AB - Gene complementation is a powerful tool for genetic selection and protein functional studies but typically requires extensive screening for complementary components. Here we engineered a platform for gene complementation based on RNA end-joining (REJ) that precisely and efficiently splices separate RNA units into functional coding mRNAs. REJ is mediated by short structured modular RNA segments of ~250bp that: (a) promote RNA::RNA interaction, (b) coopt the intrinsic cell splicing machinery to facilitate RNA trans-splicing, (c) minimizes translation of protein fragments from un-spliced RNA segments, and (d) encodes scar-free protein. We demonstrate that REJ is a broadly applicable system that can reliably split nearly any gene into complementary segments regardless of protein structure. To enable the use of this system we provide a toolbox of reporters and a web-based design tool to facilitate vector design. Demonstrated REJ applications include genetic complementation, intersectional labeling of cell types, and efficient expression of large proteins.
ER -