Crispr Mutagenesis
Crispr Mutagenesis - Related Videos
10:52
Precise Phage Mutagenesis with NgTET-Assisted CRISPR-Cas Systems
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2025
Here, we present a protocol to reduce DNA modifications in bacteriophages using the NgTET enzyme, enabling efficient and scarless CRISPR-Cas mutagenesis. This method facilitates the genetic engineering of phages for applications in biotechnology and phage therapy.
07:13
CRISPR-Cas9-based Mutagenesis in the Entomopathogenic Nematode Steinernema hermaphroditum and the Maintenance of Mutant Lines
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2025
This article demonstrates CRISPR-Cas9-mediated genome engineering in Steinernema hermaphroditum, an entomopathogenic (EPN: insect-parasitic) nematode and an emerging genetic model. The described technology is useful for creating mutants, allowing for the elucidation of gene functions in nematode biology that are relevant to mutualistic and parasitic symbiosis.
08:30
Embryo Injections for CRISPR-Mediated Mutagenesis in the Ant Harpegnathos saltator
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Cited by 9 •
2021
Many characteristics of insect eusociality rely on within-colony communication and division of labor. Genetic manipulation of key regulatory genes in ant embryos via microinjection and CRISPR-mediated mutagenesis provides insights into the nature of altruistic behavior in eusocial insects.
08:19
Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis
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Cited by 8 •
2022
This paper presents the step-by-step protocols for CRISPR/Cas9 mutagenesis of the Oriental fruit fly Bactrocera dorsalis. Detailed steps provided by this standardized protocol will serve as a useful guide for generating mutant flies for functional gene studies in B. dorsalis.
02:53
CRISPR and crRNAs
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2020
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea. The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
FAQs
The repair pathway operating after the nuclease-induced break strongly influences the resulting genetic change. Error-prone nonhomologous end joining can generate mutations, whereas homology-directed repair uses a template to support precise sequence changes. Consequently, the same targeted cleavage strategy can produce either disruptive alterations or defined edits, depending on how the cell repairs the damaged DNA.
Nonhomologous end joining and homology-directed repair serve different experimental goals. Nonhomologous end joining is error-prone and can create mutations suitable for disrupting gene function. Homology-directed repair instead follows a template-guided process that can introduce a precise sequence change. This distinction lets researchers choose between mutation-based gene knockouts and more controlled variant analysis.
Guide RNA complementarity determines which DNA sequence the CRISPR-associated nuclease is directed toward. A matching sequence provides the targeting information needed for site-specific cleavage, linking the selected genomic location to the subsequent repair outcome. In genetics experiments, this targeting relationship is essential because the resulting mutation or sequence change must be associated with the gene or variant under study.
A CRISPR mutagenesis workflow begins by directing a guide RNA and nuclease to a selected complementary DNA sequence. The nuclease then creates a site-specific double-strand break, after which cellular repair generates either mutations or precise sequence changes. Researchers can interpret the resulting DNA alteration in relation to the targeted gene, regulatory question, or genetic variant.
Researchers use CRISPR mutagenesis when they need to connect a particular DNA sequence with gene function or biological traits. Error-prone repair can support gene knockouts by generating disruptive mutations, while template-guided repair can enable analysis of defined sequence variants. These applications make the approach useful for testing functional effects rather than only observing genetic associations.
In genetics, CRISPR mutagenesis provides a way to examine how targeted DNA changes influence biological traits. Its applications include gene knockouts, variant analysis, functional screens, and disease modeling. By linking an intentional sequence alteration to an observed outcome, the method helps researchers investigate gene function and genetic regulation within experimental systems.