Creation of a High-Resolution, Continually Evolvable Lineage Tracer
- 1Harvard Medical School
Abstract
Cellular diversity is required for the proper function of many tissues, including the retina, with its ~130 cell types.1-3 Previous lineage studies suggested that retinal progenitor cells (RPCs) generate cell diversity through biased cell divisions.4-9 However, the extent to which each progenitor cell contributes to diversity, and the overarching rules governing the lineages of the collection of progenitor cells, are unknown. While classic lineage tracing approaches including retroviral labeling4,6,10, dye injection11,12, and time-lapse microscopy13,14 were used to make the foundational insights regarding retinal lineages, they cannot be used to interrogate multigenerational lineages at scale. Here we describe the creation of an evolvable lineage tracing tool, "SCRIBBLE" for Sequential Combinatorial Recorder for Iterative Barcode-Based Lineage Evolution. SCRIBBLE uses prime editor to iteratively edit a barcode sequence while preserving the order of genesis of clonally related progeny. Through a series of rationally designed high-throughput screens and design optimizations, we identified novel sequences that support a high level of barcode editing and information content. Evaluation of SCRIBBLE in embryonic mouse retina recapitulated known lineages in retinal development, demonstrating that SCRIBBLE is capable of high-resolution lineage tracing in a complex tissue.
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BibTeX
@misc{delgado2026creation,
title = {{Creation of a High-Resolution, Continually Evolvable Lineage Tracer}},
author = {R. N. Delgado and Y. Si and R. E. Andersen and C. Lee and H. L. Bushnell and J. L. Doman and S. Pandey and A. A. Sousa and E. R. West and C. E. Dagotto and D. R. Liu and C. L. Cepko},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.30.754670},
url = {https://www.biorxiv.org/content/10.64898/2026.09.30.754670v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Creation of a High-Resolution, Continually Evolvable Lineage Tracer
AU - R. N. Delgado
AU - Y. Si
AU - R. E. Andersen
AU - C. Lee
AU - H. L. Bushnell
AU - J. L. Doman
AU - S. Pandey
AU - A. A. Sousa
AU - E. R. West
AU - C. E. Dagotto
AU - D. R. Liu
AU - C. L. Cepko
PY - 2026
DA - 2026/09/30
PB - bioRxiv
DO - 10.64898/2026.09.30.754670
UR - https://www.biorxiv.org/content/10.64898/2026.09.30.754670v1
AB - Cellular diversity is required for the proper function of many tissues, including the retina, with its ~130 cell types.1-3 Previous lineage studies suggested that retinal progenitor cells (RPCs) generate cell diversity through biased cell divisions.4-9 However, the extent to which each progenitor cell contributes to diversity, and the overarching rules governing the lineages of the collection of progenitor cells, are unknown. While classic lineage tracing approaches including retroviral labeling4,6,10, dye injection11,12, and time-lapse microscopy13,14 were used to make the foundational insights regarding retinal lineages, they cannot be used to interrogate multigenerational lineages at scale. Here we describe the creation of an evolvable lineage tracing tool, "SCRIBBLE" for Sequential Combinatorial Recorder for Iterative Barcode-Based Lineage Evolution. SCRIBBLE uses prime editor to iteratively edit a barcode sequence while preserving the order of genesis of clonally related progeny. Through a series of rationally designed high-throughput screens and design optimizations, we identified novel sequences that support a high level of barcode editing and information content. Evaluation of SCRIBBLE in embryonic mouse retina recapitulated known lineages in retinal development, demonstrating that SCRIBBLE is capable of high-resolution lineage tracing in a complex tissue.
ER -