Grna Cas9
Grna Cas9 - Related Videos
04:22
A Technique for Gene Editing in Natural Killer Cells Using CRISPR Cas9
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2025
This video demonstrates a technique for Cas9 ribonucleoprotein-mediated genetic modification of primary natural killer (NK) cells. A Cas9 ribonucleoprotein, consisting of a Cas9 endonuclease bound to a guide RNA (gRNA) formed by base pairing a CRISPR RNA (crRNA) and a trans-activating crRNA (tracrRNA), is introduced into primary natural killer cells via electroporation. The ribonucleoprotein targets and cleaves the host DNA at the target site, leading to gene knockout via modification of the...
10:32
A Universal Protocol for Large-scale gRNA Library Production from any DNA Source
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Cited by 2 •
2017
Methods for generating large-scale gRNA libraries should be simple, efficient and cost-effective. We describe a protocol for the production of gRNA libraries based on enzymatic digestion of target DNA. This method, CORALINA (comprehensive gRNA library generation through controlled nuclease activity) presents an alternative to costly custom oligonucleotide synthesis.
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.
09:10
Direct Gene Knock-out of Axolotl Spinal Cord Neural Stem Cells via Electroporation of CAS9 Protein-gRNA Complexes
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Cited by 6 •
2019
Presented here is a protocol to perform time- and space-restricted gene knock-out in axolotl spinal cords by injecting CAS9-gRNA complex into the spinal cord central canal followed by electroporation.
11:31
Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells
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Cited by 23 •
2016
Experimental validation of enhancer activity is best approached by loss-of-function analysis. Presented here is an efficient protocol that uses CRISPR/Cas9 mediated deletion to study allele-specific regulation of gene transcription in F1 ES cells which contain a hybrid genome (Mus musculus129 x Mus castaneus).
FAQs
The protospacer adjacent motif, or PAM, serves as a nearby DNA landmark for Cas9 activity. A complementary guide sequence alone is therefore not sufficient; the target must also lie next to a PAM so Cas9 can engage the locus and cut. This requirement helps determine which genomic sites are available for experimental targeting.
When Cas9 creates a double-strand break, cellular repair can produce different experimental outcomes. Nonhomologous end joining can disrupt the affected gene, supporting loss-of-function studies. Homology-directed repair can instead introduce a defined sequence change. The repair outcome therefore connects the same cutting event to different questions about gene function or a specified genetic alteration.
The guide RNA provides the sequence-level matching information because its sequence is complementary to the intended DNA target. Cas9 is directed to that locus when this complementarity is paired with a nearby PAM. Consequently, changing the guide sequence changes the genomic site under investigation, allowing different genes or regions to be examined.
A basic workflow begins by selecting a developmental gene or DNA sequence whose function is being studied, then using the guide and Cas9 system to perturb that target in an embryo, stem cell, or model organism. Investigators can evaluate the resulting effects on cell fate, tissue formation, or broader developmental processes, linking targeted genetic change to biological outcome.
These settings allow investigators to examine targeted gene function in different developmental contexts. Work in embryos, stem cells, and model organisms can address effects on cell fate, tissue formation, and broader developmental processes. This range makes the system useful for connecting a specific genetic perturbation with how developmental processes operate across experimental models.
By making targeted changes in genes associated with development, investigators can examine how altered gene function relates to cell fate, tissue formation, and developmental processes. The same approach can also help investigate the genetic basis of developmental disorders. Its value lies in connecting a specific sequence change with developmental consequences in embryos, stem cells, or model organisms.