Abstract
The technique of Cre-mediated DNA recombination at loxP sites has been used widely in manipulation of the genome in cultured cells and in living organisms. Local delivery of Cre recombinase protein tagged with a cell-penetrating (or permeable) peptide (Cre-CPP) has the advantage of additional spatial and temporal control when compared to genetic delivery methods. In this chapter, we describe protocols for injection-based intramuscular delivery of Cre-CPP dissolved in hydrogel to skeletal muscle and by ultrasound-guided injection to cardiac muscle in mice.
Keywords:
Cardiomyocyte; Cell-penetrating peptide; Cre recombinase; Extensor digitorum longus; Intramuscular injection; Skeletal muscle; Tibialis anterior; Ultrasound-guided injection.
MeSH terms
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Animals
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Cell-Penetrating Peptides / administration & dosage
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Cell-Penetrating Peptides / genetics*
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Cell-Penetrating Peptides / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Fibroblasts / cytology
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Fibroblasts / metabolism
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Gene Editing / methods*
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Genetic Loci
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Genome*
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Hydrogels / administration & dosage
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Hydrogels / chemistry
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Injections, Intramuscular
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Integrases / administration & dosage
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Integrases / genetics*
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Integrases / metabolism
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Mice
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Muscle, Skeletal / metabolism
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Myocardium / metabolism
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Myocytes, Cardiac / cytology
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Myocytes, Cardiac / metabolism
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Plasmids / administration & dosage
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Plasmids / chemistry
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Plasmids / metabolism
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Primary Cell Culture
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Recombinant Fusion Proteins / administration & dosage
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Recombinant Fusion Proteins / genetics*
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Recombinant Fusion Proteins / metabolism
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Recombination, Genetic*
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Ultrasonography
Substances
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Cell-Penetrating Peptides
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Hydrogels
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Recombinant Fusion Proteins
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Cre recombinase
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Integrases