Microhomology-mediated End Joining
Microhomology-mediated End Joining - Related Videos
08:22
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
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Cited by 15 •
2018
The clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) system provides a promising tool for genetic engineering, and opens up the possibility of targeted integration of transgenes. We describe a homology-mediated end joining (HMEJ)-based strategy for efficient DNA targeted integration in vivo and targeted gene therapies using CRISPR/Cas9.
04:13
CRISPR Concatemer-Mediated Multiple Gene Knockout: A Technique to Simultaneously Knockout Multiple Genes by Non-Homologous End-Joining Pathway in Mouse Intestinal Cells
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2025
This video describes a gene knockout technique using a CRISPR-concatemer to simultaneously knock out multiple genes in cultured mouse intestinal organoid cells. This method is used to knock out a diseased gene and to elucidate the function of a gene and its paralogues.
06:10
Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates
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2025
A detailed protocol is provided for using CRISPR/Cas9 technology to achieve highly efficient targeted knock-in of large, multicistronic constructs in primary human T cells via the homology-mediated end joining (HMEJ) DNA repair pathway. T cells engineered with this cGMP-adaptable protocol maintain excellent cell expansion, cytotoxicity, and cytokine production.
11:35
Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Cited by 6 •
2017
Recent advances in the ability to genetically manipulate somatic cell lines hold great potential for basic and applied research. Here, we present two approaches for CRISPR/Cas9 generated knockout production and screening in mammalian cell lines, with and without the use of selectable markers.
02:27
Nonsense-mediated mRNA Decay
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2020
The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs. Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
FAQs
The pathway becomes error-prone because repair does not simply restore the original chromosome ends. Nucleases first resect damaged DNA, exposing short matching regions. These microhomologies align, DNA synthesis extends the aligned ends, and ligation completes repair. This sequence of events can produce deletions or other changes at the original break site.
DNA polymerase theta is a key enzyme associated with MMEJ, making it an important factor for mechanistic and therapeutic studies. Examining its activity can help researchers determine how tumor cells repair broken DNA and whether cancers with defects in other repair pathways have a selective vulnerability that could be targeted.
The comparison helps researchers investigate whether tumors lacking effective activity in another repair pathway become more reliant on MMEJ. If that reliance is important for survival, disrupting MMEJ-related factors such as DNA polymerase theta could selectively affect those cancer cells while clarifying how repair pathway defects shape tumor biology.
Because repair through MMEJ can introduce deletions and other sequence changes, repeated use of the pathway may alter chromosome sequences over time. In cancer research, these repair-associated changes are relevant to genomic instability and may support tumor evolution by increasing the genetic variation within developing tumor cells.
Studies can evaluate the relationship between MMEJ activity, DNA sequence changes at break sites, genomic instability, tumor evolution, and treatment response. These outcomes help connect a molecular repair process with broader cancer behavior and may reveal whether altered repair activity contributes to resistance against DNA-damaging treatments.
Targeting MMEJ becomes especially relevant when researchers study cancers carrying defects in other DNA repair pathways. Investigating DNA polymerase theta and related activity may identify tumors that depend on this error-prone repair route. Such findings could guide selective treatment strategies and help explain differences in sensitivity to DNA-damaging therapies.