ArticleMolecular therapy. Nucleic acids2018
Development of a Reporter System to Explore MMEJ in the Context of Replacing Large Genomic Fragments.
Article in Molecular therapy. Nucleic acids, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 19 citations in OpenAlex.
- CRISPR/Cas9-Based Genome Editing: Understanding Differences in DNA Repair Pathways, Profiles, and Outcomes.International journal of molecular sciences · 2026Review
- Influence of PARP1 on CRISPR/Cas9 induced double strand break repair in proliferating cells.Computational and structural biotechnology journal · 2025Article
- Targeting DNA repair pathways with B02 and Nocodazole small molecules to improve CRIS-PITCh mediated cassette integration in CHO-K1 cells.Scientific reports · 2023Article
- CRISPR/Cas9 System: A Potential Tool for Genetic Improvement in Floricultural Crops.Molecular biotechnology · 2022Review
- Translational potential of base-editing tools for gene therapy of monogenic diseases.Frontiers in bioengineering and biotechnology · 2022Review
- Genome Editing for β-Hemoglobinopathies: Advances and Challenges.Journal of clinical medicine · 2021Review
- Using Transcriptomic Analysis to Assess Double-Strand Break Repair Activity: Towards Precise in vivo Genome Editing.International journal of molecular sciences · 2020Article
- Microhomologies are prevalent at Cas9-induced larger deletions.Nucleic acids research · 2019Article
- LION: a simple and rapid method to achieve CRISPR gene editing.Cellular and molecular life sciences : CMLS · 2019Article
- Understanding the diversity of genetic outcomes from CRISPR-Cas generated homology-directed repair.Communications biology · 2019Article
Corrections and comments
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Authors and funding
12 authors at 2 institutions in 2 countries.
Funding
Abstract
Common genome-editing strategies are either based on non-homologous end joining (NHEJ) or, in the presence of a template DNA, based on homologous recombination with long (homology-directed repair [HDR]) or short (microhomology-mediated end joining [MMEJ]) homologous sequences. In the current study, we aim to develop a model system to test the activity of MMEJ after CRISPR/Cas9-mediated cleavage in cell culture. Following successful proof of concept in an episomally based reporter system, we tested template plasmids containing a promoter-less luciferase gene flanked by microhomologous sequences (mhs) of different length (5, 10, 15, 20, 30, and 50 bp) that are complementary to the mouse retinitis pigmentosa GTPase regulator (RPGR)-ORF15, which is under the control of a CMV promoter stably integrated into a HEK293 cell line. Luciferase signal appearance represented successful recombination events and was highest when the mhs were 5 bp long, while longer mhs revealed lower luciferase signal. In addition, presence of Csy4 RNase was shown to increase luciferase signaling. The luciferase reporter system is a valuable tool to study the input of the different DNA repair mechanisms in the replacement of large DNA sequences by mhs.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.