Nanopore sequencing identifies a new Tyr::CreERT2 allele circulating in existing mouse stocks
- 1Lancaster University
Abstract
The Tyr::CreERT2 transgenic mouse lines are essential tools for conditional gene manipulation in melanocytes and are widely used in melanoma research. Two independent lines are in common use: the Bosenberg line and the Larue line. Precise knowledge of transgene integration sites is critical for designing complex genetic crosses, yet the integration sites for these lines have only recently been characterised by whole genome sequencing. Here we report that a Tyr::CreERT2 mouse stock routinely used in BrafCA;Ptenflox melanoma models carries a previously undescribed integration on Chromosome 1, distinct from the previously reported Chromosome 2 integration. Using long-read nanopore sequencing, we mapped the integration to an intergenic locus between Alppl2 and Alpi, revealing a 2,430 bp genomic deletion at the insertion site. We developed position-specific junction PCR and qPCR assays to genotype this allele and confirmed that offspring are born at Mendelian ratios. Given that this stock is associated with elevated spontaneous melanoma penetrance, accurate characterisation of this allele has direct implications for the many laboratories employing this widely distributed melanoma model.
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BibTeX
@misc{pomfret2026nanopore,
title = {{Nanopore sequencing identifies a new Tyr::CreERT2 allele circulating in existing mouse stocks}},
author = {L. Pomfret and A. Nugawela and E. Wilkinson and B. B.-J. Shih and R. Mort},
howpublished = {bioRxiv},
year = {2026},
month = jul,
doi = {10.64898/2026.07.29.741227},
url = {https://www.biorxiv.org/content/10.64898/2026.07.29.741227v2},
note = {CC BY},
}
RIS
TY - UNPB
TI - Nanopore sequencing identifies a new Tyr::CreERT2 allele circulating in existing mouse stocks
AU - L. Pomfret
AU - A. Nugawela
AU - E. Wilkinson
AU - B. B.-J. Shih
AU - R. Mort
PY - 2026
DA - 2026/07/29
PB - bioRxiv
DO - 10.64898/2026.07.29.741227
UR - https://www.biorxiv.org/content/10.64898/2026.07.29.741227v2
AB - The Tyr::CreERT2 transgenic mouse lines are essential tools for conditional gene manipulation in melanocytes and are widely used in melanoma research. Two independent lines are in common use: the Bosenberg line and the Larue line. Precise knowledge of transgene integration sites is critical for designing complex genetic crosses, yet the integration sites for these lines have only recently been characterised by whole genome sequencing. Here we report that a Tyr::CreERT2 mouse stock routinely used in BrafCA;Ptenflox melanoma models carries a previously undescribed integration on Chromosome 1, distinct from the previously reported Chromosome 2 integration. Using long-read nanopore sequencing, we mapped the integration to an intergenic locus between Alppl2 and Alpi, revealing a 2,430 bp genomic deletion at the insertion site. We developed position-specific junction PCR and qPCR assays to genotype this allele and confirmed that offspring are born at Mendelian ratios. Given that this stock is associated with elevated spontaneous melanoma penetrance, accurate characterisation of this allele has direct implications for the many laboratories employing this widely distributed melanoma model.
ER -