Adenine Base Editing
Adenine Base 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...
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.
08:31
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...
09:56
Improved Genome Editing via Oviductal Nucleic Acids Delivery-based In Vivo Electroporation Technique for Knockout Mice Generation
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2025
The efficiency of the Improved Genome Editing via Oviductal Nucleic Acids Delivery (I-GONAD) method is comparable to traditional microinjection, which requires zygote collection from donor females and transfer to pseudo-pregnant females. This protocol demonstrates its effectiveness by introducing CRISPR/Cas9-induced mutations into the ROSA26 locus on chromosome 6.
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.
FAQs
Each component contributes a different step. The guide RNA provides sequence complementarity, positioning the engineered complex at the intended DNA region. Cas9 nickase creates a nick rather than a conventional double-strand break, while the adenine deaminase chemically converts a targeted adenine into inosine. Together, these components enable a defined base-pair change through coordinated targeting, modification, and DNA processing.
Inosine serves as the chemical intermediate that allows the original adenine-containing information to be interpreted differently. During cellular DNA repair and replication, inosine is interpreted as guanine. This processing changes the informational outcome at the edited position, while nicking the opposite DNA strand promotes fixation of the resulting G•C base pair.
The key distinction is the type of DNA alteration initiated by the editing complex. Instead of creating a conventional double-strand DNA break, the system uses adenine deamination together with a nick in the opposite strand. This strategy directs a specific A•T-to-G•C substitution through base conversion and cellular processing rather than through repair of a full double-strand break.
A typical workflow begins by selecting a guide RNA complementary to the sequence containing the intended target adenine. The guide RNA directs the engineered Cas9 nickase–adenine deaminase complex to that region, where deamination produces inosine. Cellular repair and replication then interpret the intermediate as guanine, and nicking the opposite strand helps fix the edited base pair.
In functional genomics, targeted base changes can help investigators examine how individual sequence differences affect biological function. By introducing an A•T-to-G•C substitution at a selected genomic site, researchers can investigate the consequences of a point mutation or altered sequence context. The resulting edit supports studies connecting specific DNA changes with gene activity or cellular traits.
Adenine Base Editing can support disease-model development by introducing selected point mutations into genomic DNA without initiating a conventional double-strand break. This allows researchers to create models centered on particular sequence changes, including variants relevant to disease. Such models provide a biological setting for examining how a mutation influences gene function and disease-associated processes.
The technique is relevant when a research question depends on changing a defined adenine-containing base pair. Researchers can investigate pathogenic variants by testing whether a targeted sequence change alters biological function, or examine gene regulation by modifying a point mutation associated with regulatory behavior. These applications connect precise sequence editing with the study of disease mechanisms and gene control.