Genomic Knock-in
Genomic Knock-in - 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.
08:31
Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
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Cited by 2 •
2013
Single Virus Genomics (SVG) is a method to isolate and amplify the genomes of single virons. Viral suspensions of a mixed assemblage are sorted using flow cytometry onto a microscope slide with discrete wells containing agarose, thereby capturing the virion and reducing genome shearing during downstream processing. Whole genome amplification is achieved using multiple displacement amplification (MDA) resulting in genomic material that is suitable for sequencing.
08:31
Genome Editing
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2023
A well-established technique for modifying specific sequences in the genome is gene targeting by homologous recombination, but this method can be laborious and only works in certain organisms. Recent advances have led to the development of “genome editing”, which works by inducing double-strand breaks in DNA using engineered nuclease enzymes guided to target genomic sites by either proteins or RNAs that recognize specific sequences. When a cell attempts to repair this damage, mutations can be...
09:17
Generation of Maternal Mutants Using zpc:cas9 Knock-in Zebrafish
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Cited by 1 •
2025
Here, we describe a protocol for generating maternal mutant that couples a stable zpc:cas9 knock-in line with Tol2-mediated delivery of sgRNA expression cassettes.
08:04
Determination of Sialic Acids in Liver and Milk Samples of Wild-type and CMAH Knock-out Mice.
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Cited by 7 •
2017
We describe a HPLC-based method for the determination of N-acetylneuraminic acid and N-glycolylenuraminic acid in mouse liver and milk.
FAQs
Successful editing depends on coordinating two elements with different roles. A targeted nuclease, such as CRISPR-Cas9, creates a DNA break at the selected genomic site. The donor template then provides the intended sequence, which the cell can use during homology-directed repair. This coordination links the chosen genomic location to the specific mutation, reporter, or functional sequence being introduced.
Homology-directed repair matters because it connects the donor template to the nuclease-generated break at a defined locus. Repair through this pathway can place the intended DNA sequence within its surrounding genomic context rather than treating the sequence as an independent genetic element. That relationship supports controlled analysis of gene variants, regulatory elements, and reporter behavior.
The inserted sequence determines the biological question the engineered system can address. A point mutation can test the effect of a specific genetic change, while a fluorescent reporter can support studies of protein localization. Regulatory elements can be examined for their influence on gene control, and functional gene variants can help investigate altered gene activity in cells or model organisms.
A typical workflow begins by selecting the genomic site and defining the sequence to be introduced. Researchers then use a targeted nuclease to create a site-specific DNA break and provide a donor template carrying the desired DNA. The resulting homology-directed repair is examined in the engineered cells or model organism to study the intended genetic change.
This approach is useful when researchers need to examine a defined sequence change while retaining the surrounding genomic context. It can support studies of gene regulation, disease mechanisms, protein localization, and therapeutic strategies. Because the introduced sequence is selected in advance, the resulting system can connect a specific genetic alteration with a measurable cellular or organismal outcome.
Researchers can introduce functional gene variants or point mutations into cells and model organisms to investigate how particular genetic changes affect biology. These engineered systems provide a context for examining disease-related mechanisms and for exploring therapeutic strategies. Reporter sequences and regulatory elements can add information about gene activity or protein behavior during those investigations.