Homology Directed Repair
Homology Directed Repair - Related Videos
05:12
All-in-One CRISPR Genome Editing: A Method for Homology Directed Repair-Based Gene Knock-In in Cultured Cells Using CRISPR-Cas9 System
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2025
In this video, we demonstrate all-in-one CRISPR-Cas9 based genome editing in cultured cells where Cas9 and sgRNA are provided as a single plasmid construct to the cells. The CRISPR-Cas9 system and desired gene to be inserted was introduced in cells through electroporation technique to facilitate successful gene editing.
02:31
Homologous Recombination
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2020
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
06:34
Zinc Finger Nuclease-Based Genome Editing: A Technique for Modifying Genome in Human Pluripotent Stem Cells by Double-Stranded Homology Dependant Repair Mechanism
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2025
This video demonstrates a technique of zinc-finger nuclease-based genome editing in human pluripotent stem cells (hPSCs). As hPSCs can potentially be differentiated into various cell types, this technology is helpful for in vitro pathological and pharmacological studies in a patient-specific manner.
14:23
Recombineering Homologous Recombination Constructs in Drosophila
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Cited by 9 •
2013
Homologous recombination techniques greatly advance Drosophila genetics by enabling the creation of molecularly precise mutations. The recent adoption of recombineering allows one to manipulate large pieces of DNA and transform them into Drosophila6. The methods presented here combine these techniques to rapidly generate large homologous recombination vectors.
08:53
Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1
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Cited by 15 •
2011
We provide a method for testing BRCA1 variants in a tissue culture based assay for homologous recombination repair of DNA damage by depleting endogenous BRCA1 protein from a cell using RNAi and replacing it with a BRCA1 point mutant that contains a coding change.
FAQs
After a double-strand break, the exposed single-stranded DNA ends search for a matching or nearly matching template. One end invades the template and pairs with its corresponding sequence, allowing DNA synthesis to copy information across the damaged region. The repaired strands are then resolved, restoring sequence continuity while preserving information from the chosen template.
Template matching supplies the sequence information needed to restore the damaged DNA correctly. A matching or nearly matching template provides a guide for DNA synthesis, whereas the template’s designed sequence can also specify an intended change. This principle allows HDR to support both mutation correction and the targeted insertion of defined DNA sequences in genetic studies.
HDR efficiency depends on several interacting conditions rather than on the DNA break alone. The cell-cycle state can affect whether the pathway operates efficiently, while repair-protein activity influences strand processing, template invasion, synthesis, and resolution. Template design also matters because it determines how effectively the damaged sequence can use the available genetic information for repair.
The workflow begins when the break exposes single-stranded DNA ends. Those ends locate a homologous template, invade the matching region, and use it to guide DNA synthesis. The newly restored strands must then be resolved to complete repair. In experimental genetics, researchers additionally consider the desired template sequence and the cellular conditions that support efficient repair.
Researchers use HDR when they need a defined genetic outcome rather than only restoration of damaged DNA. The pathway can support accurate correction of mutations, insertion of selected DNA sequences, and construction of engineered cell or animal models. These applications make it useful for studying gene function, modeling genetic conditions, and investigating disease-related changes.
HDR-based experiments can create cells or animals carrying a precise correction, mutation, or inserted sequence. Comparing these engineered systems with appropriate genetic backgrounds can help researchers examine how a defined DNA change affects biological function and disease-related processes. The approach therefore links a specific sequence alteration to experimental models used in genetics and disease research.