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Cre-mediated, loxP independent sequential recombination of a tripartite transcriptional stop cassette allows for partial read-through transcription.
Journal article

Cre-mediated, loxP independent sequential recombination of a tripartite transcriptional stop cassette allows for partial read-through transcription.

  • Bapst AM Institute of Physiology, University of Zürich, CH-8057 Zürich, Switzerland.
  • Dahl SL Institute of Physiology, University of Zürich, CH-8057 Zürich, Switzerland; National Centre of Competence in Research "Kidney.CH", Switzerland.
  • Knöpfel T Institute of Physiology, University of Zürich, CH-8057 Zürich, Switzerland.
  • Wenger RH Institute of Physiology, University of Zürich, CH-8057 Zürich, Switzerland; National Centre of Competence in Research "Kidney.CH", Switzerland. Electronic address: roland.wenger@access.uzh.ch.
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  • 2020-04-29
Published in:
  • Biochimica et biophysica acta. Gene regulatory mechanisms. - 2020
Cre profiling
DNA recombination
Lineage tracing
Poly(A) signal
loxP
Animals
CHO Cells
Cricetulus
Female
Gene Expression
Genes, Reporter
Genetic Markers
Integrases
Kidney
Male
Mice
Recombination, Genetic
Transcriptome
English One of the widely used applications of the popular Cre-loxP method for targeted recombination is the permanent activation of marker genes, such as reporter genes or antibiotic resistance genes, by excision of a preceding transcriptional stop signal. The STOP cassette consists of three identical SV40-derived poly(A) signal repeats and is flanked by two loxP sites. We found that in addition to complete loxP-mediated recombination, limiting levels of the Cre recombinase also cause incomplete recombination of the STOP cassette. Partial recombination leads to the loss of only one or two of the three identical poly(A) repeats with recombination breakpoints always precisely matching the end/start of each poly(A) signal repeat without any relevant similarity to the canonical or known cryptic loxP sequences, suggesting that this type of Cre-mediated recombination is loxP-independent. Incomplete deletion of the STOP cassette results in partial read-through transcription, explaining at least some of the variability often observed in marker gene expression from an otherwise identical locus.
Language
  • English
Open access status
hybrid
Identifiers
Persistent URL
https://folia.unifr.ch/global/documents/65840
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