Mitochondrion-targeted gene editing complex, preparation method and application thereof and mitochondrion genome editing method
A gene editing and mitochondrial technology, applied in the biological field, can solve problems such as difficulty in restoring target gene function, inability to treat oxidative stress, and physiological function effects, achieving significant antioxidant capacity, improving oxidative stress levels, and highly targeted treatments. Effect
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[0048] 1. Synthesis of mitochondria-targeted cationic liposomes:
[0049] Cationic liposomes are used to synthesize ordinary liposomes (Liposomes) with phospholipids (DOTAP) and cholesterol (Chol); the liposomes are modified with the classic mitochondrial targeting agent triphenylphosphine to endow the liposomes with mitochondrial targeting properties; On this basis, liposomes are covalently bonded with maleimide polyethylene glycol-FITC (MAL-PEG-FITC) micellar solution to increase its water solubility and stability, reduce immunogenicity, and simultaneously Give it a FITC fluorescent label.
[0050] 2. Construction and assembly of mitochondria-targeted gene editing complex (LCCT complex)
[0051] Use the region between 15953 and 15956 as the restriction site, and use U6 as the promoter to synthesize the corresponding crRNA;
[0052]
[0053] The double-stranded homologous template of TERT (dsDNA donor-TERT-mcherry) tagged with the reporter gene mcherry was constructed; t...
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