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Analytical Chem JoVE (Open Access) Citable · DOI

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

DOI: 10.3791/52599-v
What you'll learn
  • ✓ Generate and validate inducible telomerase knock-in mouse alleles (mTERT-ER and LSL-mTERT)
  • ✓ Reactivate telomerase in vivo and isolate telomerase-expressing neural stem cells in vitro
  • ✓ Assess telomere length changes using fluorescence in situ hybridization (FISH)
  • ✓ Evaluate telomerase reactivation effects on tissue regeneration and cancer models
Protocol

Biopharma Insights Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer.

Difficulty
advanced
Total time
~3-4 weeks (breeding/genotyping) + 1-2 weeks (induction/analysis per experiment)
Model organism
Mouse (murine inducible telomerase alleles: mTERT-ER and LSL-mTERT)
Biosafety
BSL-1

Steps

1
Reactivate telomerase in transgenic mice in vivo

Induce telomerase expression in mTERT-ER and LSL-mTERT mouse lines using appropriate induction methods (e.g., tamoxifen administration or Cre-recombinase activation) to study tissue regeneration and degeneration responses.

▶ 01:35
2
Isolate neural stem cells and activate telomerase in vitro

Extract neural stem cells from induced mice and culture them with telomerase reactivation to observe proliferation and differentiation effects in a controlled laboratory setting.

▶ 02:33
3
Perform telomere fluorescence in situ hybridization

Apply FISH methodology to visualize and quantify telomere length in samples from telomerase-reactivated and control tissues or cells.

▶ 05:07
4
Analyze telomerase reactivation effects on tissue phenotypes

Compare outcomes between induced and control samples to assess impacts on tissue regeneration, cellular senescence, and tumorigenesis markers.

▶ 06:37
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