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Updated: Jun 2, 2026

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
10:07

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

Targeted correction of point mutations in the low density lipoprotein receptor gene mediated by single-stranded

Fang-Lin Zhang1, Wei Dong, Xiao-Yan Wu

  • 1Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029, P.R. China. zhangfl05@gmail.com.

Molecular Medicine Reports
|April 9, 2011
PubMed

Insights

Researchers explored correcting low-density lipoprotein receptor (LDLR) gene mutations using single-stranded oligonucleotides (SSOs) in mice. Antisense SSOs successfully repaired the LDLR gene mutation in vivo, offering a potential treatment for familial hypercholesterolemia (FH).

Area of Science:

  • Molecular Biology
  • Genetics
  • Gene Therapy

Background:

  • Familial hypercholesterolemia (FH) is primarily caused by mutations in the low-density lipoprotein receptor (LDLR) gene.
  • Single-stranded oligonucleotides (SSOs) offer sequence-specific gene alteration capabilities.
  • In vivo gene repair strategies are crucial for treating genetic disorders.

Purpose of the Study:

  • To investigate the efficacy of single-stranded oligonucleotides (SSOs) in repairing LDLR gene point mutations in vivo.
  • To evaluate the potential of SSO-mediated gene repair as a therapeutic strategy for familial hypercholesterolemia (FH).

Main Methods:

  • A construct containing the mutated LDLR gene fused to a luciferase reporter (p660-LDLR-luc) was introduced into mouse liver via hydrodynamic gene transfer.
  • Mice were injected with various SSOs complexed with polyethylenimine.
  • Repair efficiency was assessed by measuring firefly luciferase activity, restriction fragment length polymorphism, and direct sequencing.

Main Results:

  • Antisense SSOs (anti-SSOs) demonstrated successful correction of the LDLR nonsense mutation (c660x) in mouse liver in vivo.
  • Luciferase activity assays confirmed the functional repair of the gene.
  • Molecular analyses confirmed the sequence-specific correction of the LDLR mutation.

Conclusions:

  • This study provides initial evidence for successful in vivo correction of LDLR gene point mutations using anti-SSO targeted repair.
  • SSO-mediated gene repair represents a promising potential therapeutic strategy for familial hypercholesterolemia (FH).

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