# High resolution neuronal lineage tracing

> **NIH NIH R21** · NEW YORK UNIVERSITY · 2020 · $431,454

## Abstract

Project Summary
How neuronal progenitor cells produce an enormous diversity of neuronal and glial cell types is a fundamental
topic that remains largely unresolved. Drosophila has been a pivotal model system to study these complex
questions of neurogenesis and research in its nervous system has contributed to several important concepts
that apply to mammals. These include temporal and spatial patterning, cell death, neural and/or glial specification
and asymmetric cell division. Nevertheless, the full resolution of its neuronal lineages remains elusive. In this
proposal we will take advantage of powerful genetic tools derived from synthetic biology to reconstruct the entire
neuronal lineage of multiple brain structures at single cell resolution. We will develop transgenic flies in which
lineages can be autonomously recorded and analyzed through single cell transcriptomics. We hypothesize that
our knowledge on the structure and molecular nature of adult brain development will provide us with a unique
advantage to not only reconstruct the entire lineage tree of the Drosophila brain, but also to form new hypotheses
on how each of the brain structures form very faithfully and uniformly from precursor cells.
Aim 1 Development of a progressive lineage recorder in Drosophila. There are currently no established
CRISPR-based lineage methods in the Drosophila nervous system. We will adapt GESTALT in Drosophila to
`scar' the DNA during lineage progression and progressively record this lineage through development. We will
use genetic tools to gain spatio-temporal control over our lineage measurements and use in silico modeling of
the barcode structure to optimize the activity of the system. We will generate flies with enough target sites to
capture the entire neuronal diversity generated during neuronal development. We will empirically evaluate
different versions of the technology and select the best one for single cell lineage tracing.
Aim 2 Defining neuronal birth order and clonal relationships in the adult brain. We will lineage trace
through the neuronal development while simultaneous sequencing the transcriptome of single cells. This should
allow us to identify the different neural subtypes using our single cell atlas. We will characterize the lineage
information per cell and combine this with the published methods capable of reconstructing multi-tree lineages
to identify different lineage relationships in our data. We will build on the stereotypical mode of neuronal
development to refine this structure and reconstruct the neuronal lineages
Aim 3 Experimental validation of the reconstructed neuronal lineages. We will use our prior knowledge of
neuronal development in combination with post hoc validation to benchmark our lineage reconstruction. We will
first compare our lineage reconstruction of the local motion detectors in Drosophila to their known simple and
well-defined lineage. We will use region-specific Gal4 lines to lineage trace different subreg...

## Key facts

- **NIH application ID:** 10042321
- **Project number:** 1R21NS117972-01
- **Recipient organization:** NEW YORK UNIVERSITY
- **Principal Investigator:** Claude Desplan
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $431,454
- **Award type:** 1
- **Project period:** 2020-08-01 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10042321

## Citation

> US National Institutes of Health, RePORTER application 10042321, High resolution neuronal lineage tracing (1R21NS117972-01). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10042321. Licensed CC0.

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