Pathway Editing
Pathway Editing - Related Videos
02:23
RNA Editing
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2020
RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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...
02:58
CRISPR-Mediated Base Editing Tools: A Genome Editing Technique to Induce Targeted Base Substitution
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2025
This video explains the concept of CRISPR-mediated cytosine base editors for inducing targeted nucleotide substitution.
08:50
A Nonsequencing Approach for the Rapid Detection of RNA Editing
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2022
Rapid detection and reliable quantification of RNA editing events at a genomic scale remain challenging and currently rely on direct RNA sequencing methods. The protocol described here uses microtemperature gradient gel electrophoresis (µTGGE) as a simple, quick, and portable method of detecting RNA editing.
02:30
A CRISPR-Cas9 Technique for Gene Editing in T Cells
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2025
This video demonstrates an assay for performing gene editing in human T cells using the CRISPR-Cas9 technology. A mixture of primary CD4+ and CD8+ T cells is combined with a CRISPR-Cas9 ribonucleoprotein complex, targeting specific genes for knockout. Upon electroporation, the sgRNA guides Cas9 to the target DNA sequence, creating precise cuts. These cuts are then repaired by the cell's non-homologous end-joining mechanism, leading to gene knockout.
FAQs
Changing a selected component and examining the resulting cellular outcome helps connect that component to a specific function. If the change alters molecule production, signaling activity, or information processing, the comparison provides evidence that the targeted step contributes to that outcome. This causal perspective helps distinguish pathway-associated changes from effects occurring elsewhere in the cell.
Redirecting flux means changing how biochemical processing is distributed through a pathway by modifying selected steps rather than rebuilding the entire system. The edited component can shift processing toward or away from particular molecular products. This makes the approach useful for studying metabolic control and for designing cells whose production behavior is intentionally adjusted.
Regulatory elements can be targeted when the goal is to alter gene regulation rather than directly replace a gene product. Such an edit may change how strongly or under what conditions a pathway component contributes to cellular activity. Comparing that result with a gene-focused edit can help separate control of expression from the downstream function of the encoded component.
Researchers compare edited and unedited pathways and then examine the cellular outcome relevant to the experiment, such as signaling activity or production of a molecule. The unedited comparison establishes baseline behavior, while the edited condition shows the consequence of the targeted change. Interpreting both together helps identify pathway steps linked to the measured result.
The approach can support investigations of metabolism, gene regulation, development, and disease mechanisms, while also helping engineers create cells that produce valuable compounds. The appropriate target depends on the question: a disease study may focus on pathway changes associated with cellular dysfunction, whereas a production system may seek altered biochemical flux and a defined molecular output.
By linking selected pathway changes to measurable cellular outcomes, the approach can identify mechanisms relevant to disease and inform the design of therapies or diagnostic strategies. In engineered biology, the same reasoning supports cells with defined functions, including controlled production of valuable compounds. Its broader value comes from connecting molecular changes with outcomes that can be tested and compared.