Genomic Modification
Genomic Modification - Related Videos
03:48
Screening for Genomic Modifications: A Method to Identify CRISPR-Generated Mutants in Drosophila
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2023
This video describes a screening method to identify transgenic Drosophila flies, whose genome was modified using CRISPR-Cas9. CRISPR-Cas9 is a powerful genome-editing tool that revolutionized the way scientists can manipulate an organism's genome. In the example protocol, we will see how to identify CRISPR-generated mutants, in which an insertion of a transactivation sequence replaces the first exon of the branchless (bnl) gene, thus creating a bnl gene-specific driver line, bnl-LexA.
11:50
Genome-Wide Mapping of Histone Modifications and Transcription Factor Binding Sites in Neuroendocrine Small Cell Lung Cancer Cell Lines Using CUT&RUN
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2026
An optimized Cleavage Under Targets and Release Using Nuclease followed by next generation sequencing (CUT&RUN-seq) protocol is described for neuroendocrine small cell lung cancer cell lines. It enables genome-wide mapping of various histone modifications and transcription factor (e.g. E2F7) binding sites to investigate epigenetic and transcriptional deregulation in SCLC pathobiology.
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.
02:32
Histone Modification
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2020
The histone proteins have a flexible N-terminal tail extending out from the nucleosome. These histone tails are often subjected to post-translational modifications such as acetylation, methylation, phosphorylation, and ubiquitination. Particular combinations of these modifications form “histone codes” that influence the chromatin folding and tissue-specific gene expression. Acetylation The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...
09:10
A Fast and Quantitative Method for Post-translational Modification and Variant Enabled Mapping of Peptides to Genomes
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Cited by 3 •
2018
Here we present the proteogenomic tool PoGo and protocols for fast, quantitative, post-translational modification and variant enabled mapping of peptides identified through mass spectrometry onto reference genomes. This tool is of use to integrate and visualize proteogenomic and personal proteomic studies interfacing with orthogonal genomics data.
FAQs
CRISPR-Cas9 uses a guide RNA to recognize a matching DNA sequence and direct the Cas9 nuclease to that site. Cas9 then creates a break, and cellular repair processes can produce the intended alteration. This targeting mechanism allows researchers to investigate how a selected sequence affects gene function or an observable trait.
The outcome may involve altered gene activity and, consequently, a changed observable trait, even when the experimental goal is not simply to insert or delete a sequence. Researchers therefore assess both the genetic change and its effects. Variable expression can complicate interpretation, making it important to connect the modification with the observed biological result.
These approaches represent distinct ways to alter or study genetic information. CRISPR-Cas9 is specifically characterized by sequence-directed activity through a guide RNA and nuclease, whereas recombinant DNA technology and targeted mutagenesis are additional strategies identified for genomic modification. Researchers select among these approaches according to the biological question and the desired genetic or regulatory change.
An intended DNA change may not fully predict the final biological outcome. Unintended edits can introduce additional genetic changes, while variable expression can cause the same planned modification to produce different levels of gene activity or trait effects. These factors can obscure gene-function conclusions and affect the reliability of disease models, agricultural studies, or therapy-oriented research.
Validation should determine whether the intended genetic alteration occurred and whether the resulting gene activity or observable trait matches expectations. Researchers also need to consider unintended edits and variable expression, because a modification can be present without producing a uniform outcome. This evaluation supports stronger conclusions about gene function and helps identify limitations before broader biological interpretation.
Researchers use it to investigate gene function by changing selected genetic information and examining effects on gene activity or observable traits. This approach can connect a particular sequence with a biological outcome and support disease modeling. Interpretation still requires validation, since unintended edits or variable expression may produce effects that do not arise from the intended modification alone.
The overview identifies disease modeling, improvement of agricultural traits, and development of potential therapies as major applications. In each setting, the desired change must be linked to a meaningful biological outcome, while unintended edits and variable expression remain important constraints. For modifications that can persist across generations, researchers must also consider how effects may change over time.
A modification may influence not only the organism or cells examined initially but also later generations. This makes generational outcomes relevant when researchers evaluate the durability and consequences of a genetic change. Considering these effects is especially important when interpreting experimental results or assessing potential applications, because short-term observations may not capture the full outcome of the modification.