# Clonal Analysis of the Cranial Neural Crest using RIA Viruses

> **NIH NIH F31** · CALIFORNIA INSTITUTE OF TECHNOLOGY · 2020 · $46,320

## Abstract

Project Summary
 The neural crest is a population of cells that are critical for vertebrate embryonic
development. These cells are known for their migratory behavior, and contribution to a wide range
of derivatives such as the craniofacial skeleton, epidermal pigment cells, and much of the
peripheral nervous system. This proposal tackles a fundamental question in developmental
biology: whether individual cranial neural crest cells in vivo are multipotent, and retain the ability
to differentiate into numerous fates, or alternatively, if they represent a population of fate-restricted
cells, each confined to a unique cellular fate. To tackle this question, replication incompetent
avian retroviruses encoding different fluorescent fluorophores will be used to label neural
fold cells and perform clonal analyses to examine the developmental potential, movement
and morphogenesis of individual or small populations of cranial neural crest cells in vivo.
Experiments will be performed on avian embryos because of several advantages. Chick embryos
are easily accessible to retroviral infection and experimental perturbation at early stages of
development, allowing temporally and spatially controlled manipulation. Birds like humans are
amniotes but, unlike mice, develop outside the mother, making them more accessible at early
stage, while developing in a manner that is morphologically nearly identical to that of human
embryos at comparable stages.
Aim 1: Retrovirally mediated clonal analysis of the chick cranial neural crest: The cranial
neural tube of chick embryos will be infected with limiting dilutions of replication incompetent
avian retroviruses that encode fluorophores. Four different viruses reflecting different colors will
be used per embryo and clonality will be established by visual observation a few hours after
infection. The fate of clonally related cells will then be tracked using antibody staining as well as
a novel method of single molecule fluorescent in situ hybridization that allows simultaneous
analysis of up to 35 transcripts in single cells.
Aim 2: Analysis of migratory interactions between clonally related cells: The migratory
behavior of clonally related cells will be examined in whole mount, using in ovo imaging, as well
as in slice tissue sections to visualize interactions between sister cells, cousin cells and unrelated
neighbors. Once normal migratory patterns and cell interactions are established, the effects of
perturbing cell-cell and cell-substrate interactions in individual clones of migratory cells will be
examined within an otherwise normal environment.

## Key facts

- **NIH application ID:** 9872140
- **Project number:** 5F31DE027583-03
- **Recipient organization:** CALIFORNIA INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** Alison Koontz
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $46,320
- **Award type:** 5
- **Project period:** 2018-04-01 → 2021-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9872140

## Citation

> US National Institutes of Health, RePORTER application 9872140, Clonal Analysis of the Cranial Neural Crest using RIA Viruses (5F31DE027583-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9872140. Licensed CC0.

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