Zebrafish Germline Edits
Zebrafish Germline Edits - Related Videos
05:55
Screening Sperm for the Rapid Isolation of Germline Edits in Zebrafish
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Cited by 4 •
2023
CRISPR-Cas technologies have revolutionized the field of genome editing. However, finding and isolating the desired germline edit remains a major bottleneck. Therefore, this protocol describes a robust method for quickly screening F0 CRISPR-injected zebrafish sperm for germline edits using standard PCR, restriction digest, and gel electrophoresis techniques.
03:59
Gonad Microinjection: A Method of Compound Delivery Directly into the Germline of C. elegans
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2023
This video describes microinjection, a common method of creating transgenic C. elegans strains. In the example, we will see microinjection used to introduce a ribonucleoprotein (RNP) complex for CRISPR-bases gene editing.
10:37
CRISPR/Cas9-mediated Endogenous Fluorescent Tagging of Germline-specific Genes in Caenorhabditis elegans
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2026
A CRISPR/Cas9 microinjection workflow for endogenous fluorescent tagging in the Caenorhabditis elegans germline to obtain homozygous knock-in lines for in vivo protein localization analysis.
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...
10:55
Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode Caenorhabditis elegans
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Cited by 11 •
2015
We describe the detailed steps of a high-throughput chemical assay in the nematode Caenorhabditis elegans used to assess germline toxicity. In this assay, disruption of germline function following chemical exposure is monitored using a fluorescent reporter specific to aneuploid embryos.
FAQs
Cas9 first cuts a selected DNA sequence in the embryo. The cell’s repair process then determines the result: repair can create a targeted mutation, or it can incorporate a designed DNA sequence. These alternatives allow researchers to investigate the consequences of disrupting a gene or adding a specific sequence, depending on the genetic question being studied.
Introducing the editing tools at the one-cell stage places the selected DNA change in an embryo during its earliest development. When the change is present in cells that contribute to reproduction, researchers can examine whether the resulting trait appears in later generations. This timing therefore connects an initial edit with multigenerational genetic analysis.
Analysis across generations helps distinguish immediate effects of an edit from traits that persist through inheritance. Researchers can investigate how a genetic change is transmitted and whether its biological consequences remain detectable in descendants. This perspective is especially useful for studying inheritance patterns, genetic variation, and effects that may not be evident in the original edited fish.
A targeted mutation changes the selected genomic site through the cell’s repair response after Cas9 cutting. Incorporating a designed sequence instead adds a planned DNA element at that location. The two outcomes support different experimental goals: one can reveal the consequences of disrupting gene function, while the other can test the effect of introducing a specified genetic sequence.
The workflow begins by selecting a DNA sequence relevant to the research question and preparing genome-editing tools such as CRISPR-Cas9. These tools are delivered to one-cell zebrafish embryos, where Cas9 cuts the selected site. Researchers then examine the resulting genetic change and follow edited fish and their descendants when inheritance is under investigation.
Zebrafish provide a transparent, rapidly developing vertebrate model, allowing researchers to observe biological effects while studying genetic changes over development. Their use in germline-editing experiments also supports analysis across generations. Together, these features help connect a defined DNA alteration with gene function, developmental outcomes, disease mechanisms, and inheritance.
These edits can be used to investigate gene function, development, disease mechanisms, and the transmission of genetic traits. Researchers may also use the resulting models to examine genetic variation or explore therapeutic strategies. Because the changes can be followed in a developing vertebrate and across generations, the approach links molecular alterations with broader biological outcomes.