How does a Morpholino work?
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. Gene knockdown is achieved by reducing the expression of a particular gene in a cell.
What is Morpholino oligo?
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.
How long should the target specific sequence of the oligo be?
about 25-bases
Importantly, the oligo must have a base sequence that has very little self-complementarity so that it does not dimerize. Otherwise, the optimal Morpholino oligo will be about 25-bases long and have a high enough GC content (e.g. 40-60%) so that it has a high target affinity.
How do oligonucleotides work?
Oligonucleotides readily bind, in a sequence-specific manner, to their respective complementary oligonucleotides, DNA, or RNA to form duplexes or, less often, hybrids of a higher order. This basic property serves as a foundation for the use of oligonucleotides as probes for detecting specific sequences of DNA or RNA.
How do you knock down a gene?
In research laboratories, genes have traditionally been knocked down using small interfering RNA (siRNA) or short hairpin RNA (shRNA). These methods are still useful, but newer options using catalytically dead Cas9 (dCas9) or Cas13 proteins are also available. These CRISPR-based methods can offer advantages.
What are morpholino oligonucleotides useful 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.
Why are primers called oligonucleotides?
The term oligonucleotide is derived from the Greek “oligo,” which means few or small. Oligonucleotides made up of 2′-deoxyribonucleotides are the molecules used in polymerase chain reaction (PCR). These are referred to as primers and are used to massively amplify a small amount of DNA.
Are oligonucleotides primers?
For most uses, oligonucleotides are designed to base-pair with a strand of DNA or RNA. The most common use for oligonucleotides is as primers for PCR (polymerase chain reaction). Primers are designed with at least part of their sequence complementary to the 5′ end of the sequence targeted for amplification.
Are primers oligonucleotides?
How do oligonucleotides work in PCR?
They are built on an oligo building block called a phosphoramidite. During these cycles of adding one nucleotide or base to another, the chain grows in the 3′ to 5′ direction. Desalting an oligo removes the salts used in the synthesis process. After this step, the oligo is ready to use for applications like PCR.
