What are the two molecular mechanisms which Morpholino use to prevent gene expression?
Morpholinos are typically used to block translation of mRNA and to block splicing of pre‐mRNA, though they can block other interactions between biological macromolecules and RNA.
What are Morpholino oligos?
Morpholino oligos are advanced tools for blocking sites on RNA to obstruct cellular processes. A Morpholino oligo specifically binds to its selected target site to block access of cell components to that target site. Like all gene knockdown reagents, Morpholinos must be actively delivered into most cells.
Why are Morpholinos used in zebrafish?
These molecules have been applied to studies in several model organisms, including mice, zebrafish, frogs and sea urchins. Morpholinos are in development as pharmaceutical therapeutics targeted against pathogenic organisms such as bacteria or viruses and genetic diseases.
Why Morpholino?
As steric blocking oligos, Morpholinos can be used not only to block translation but also to alter mRNA splicing, to bind miRNAs or to block binding of miRNA or regulatory proteins to RNA targets. Binding of a splice-regulatory protein can be prevented by protecting its binding site with a Morpholino oligo.
What is a morpholino and what is its effect on gene expression?
Morpholinos are nucleotide analogs that recognize and bind short sequences (about 25 nucleotides) at the transcription start site or at splice sites of pre-mRNAs, and thus block the translation or proper splicing of the mRNA (Summerton & Weller, 1997).
What is Vivo morpholino?
Vivo-Morpholinos* are the knockdown, exon-skipping or miRNA blocking reagent of choice for in vivo experiments. A Vivo-Morpholino is comprised of a Morpholino oligo with a unique covalently linked delivery moiety, which is comprised of an octa-guanidine dendrimer.
What are morpholino oligonucleotides useful for?
Abstract. Caged morpholino oligonucleotides (cMOs) are useful research tools in developmental biology because they allow spatiotemporal control of gene expression in whole organisms.
What is the difference between knockout and knockdown?
The key difference between gene knockout and knockdown is that the gene knockout is a technique where the gene of interest is completely removed (inoperative state) to study of functions of the gene while gene knockdown is another technique where the gene of interest is silenced to investigate the role of the …
Can you use Morpholinos in mice?
In the mouse model, it has been shown that intravenous (IV) and intraperitoneal (IP) administration of vivo-morpholinos were equally efficacious, and recent studies have shown success with direct injection in target tissue (17-19).
Why does a target RNA morpholino hybrid not promote mRNA degradation?
Because the RNA is degraded, any sequence within the coding region of the target gene has the potential to be a useful antisense site. Morpholinos, by contrast, form RNA-morpholino hybrids that are not substrates for RNase H, and thus the mRNA is not degraded.
Are there any drawbacks to using Morpholinos?
Morpholinos are precise and efficient at interfering with genes or miRNA function, but have several potential drawbacks. They are normally injected into the yolk and affect every cell in the embryo, so may compromise normal development if the disrupted protein is required ubiquitously.
How does Morpholino reduce the dose required for a knockdown?
The dose required for a knockdown can be reduced by coinjection of several Morpholino oligos targeting the same mRNA, which is an effective strategy for reducing or eliminating dose-dependent off-target RNA interactions.
What can Morpholino oligonucleotides be used for?
Morpholinos are synthetic antisense oligonucleotides (around 25 nucleotides) designed to bind and block the translation initiation complex of messenger RNA (mRNA) sequences. This technology has been used to test the role of specific genes by transient blocking, particularly during development.
How are Morpholinos used to reverse gene function?
Morpholino. Morpholinos block access of other molecules to small (~25 base) specific sequences of the base-pairing surfaces of ribonucleic acid (RNA). Morpholinos are used as research tools for reverse genetics by knocking down gene function.