What is Morpholino compound?
Morpholinos are synthetic molecules that are the product of a redesign of natural nucleic acid structure. Usually 25 bases in length, they bind to complementary sequences of RNA or single-stranded DNA by standard nucleic acid base-pairing.
How do Morpholinos work?
Morpholinos. 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.
What is Vivo morpholino?
A Vivo-Morpholino is comprised of a Morpholino oligo with a unique covalently linked delivery moiety, which is comprised of an octa-guanidine dendrimer. It uses the active component of arginine rich delivery peptides (the guanidinium group) with improved stability and reduced cost.
What are morpholino oligonucleotides?
Morpholino oligonucleotides are nonionic DNA analogs available from Gene Tools LLC [10,11]. They possess altered backbone linkages compared with DNA or RNA (Figure 1). In spite of their altered backbone, morpholinos bind to complementary nucleic acid sequences by Watson-Crick base-pairing.
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.
How do you store Morpholino?
GENE TOOLS recommends storing a Morpholino solution at 1 mM in a tightly- sealed vial at room temperature. Storing stocks at higher concentrations can lead to variability in results.
How do you overexpress a gene?
There are four main methods for introducing foreign DNA into cells: electric shock, calcium phosphate method, liposome-mediated and virus-mediated. For many ordinary cell lines, the methods of transient transfection were mostly liposome-mediated.
What is knockout experiment?
A knockout, as related to genomics, refers to the use of genetic engineering to inactivate or remove one or more specific genes from an organism. Scientists create knockout organisms to study the impact of removing a gene from an organism, which often allows them to then learn something about that gene’s function.
How do you store Morpholinos?
Why are Morpholinos used in zebrafish?
Morpholino oligonucleotides (MOs) are the most widely used anti-sense knockdown tools in the zebrafish (Danio rerio) community. MOs have been used to accelerate gene discovery through large-scale screening,1,2 to probe candidate gene function,3 and to verify mutant phenotypes.
How do you overexpress cells?
Popular Answers (1)
- clone your gene of interest into an appropriate vector under control of the appropriate promoter.
- transfect or transduce your cells of interest using the appropriate protocol.
- begin antibiotic selection to enrich for stably integrated clones.
Can CRISPR be used to overexpress genes?
CRISPR activation (CRISPRa) is an optimized method for specific gene overexpression. CRISPRa uses an inactivated CRISPR-Cas9 system (dCas9) to upregulate target genes within their native context. This method offers many advantages over more traditional gene overexpression techniques, such as cDNA and ORF.
What is CRISPR knockdown?
A knockdown is essentially a gene that had its expression reduced, instead of being stopped altogether (as in the case of a knockout). Knockdown genes can be obtained either through genetic manipulation, through CRISPR, or with reagents such as RNA oligonucleotide or a short DNA.
What is knockout technology?
A gene knock-out (KO) is a genetic technique supplemented with biotechnological tool, in which an organism is engineered to carry genes that have been made inoperative.
How can I overexpress a gene?
How do you overexpress a gene using Crispr?
For example, if you are interested in overexpressing a single gene for a single experimental purpose, you can do so simply by packaging your gene in a lentivirus and transducing your cells. Where CRISPRa really shines is in its application to genome-wide pooled overexpression screens.
How do I overexpress a gene?
How do you overexpress proteins?
Once you have plasmid with the clone, you need to transfect cells. Hek293 are easy to transfect, I use lipofectamin or prime jet reagent. After this you need to do antibiotic selection in order to generate a new cell line that will over express the protein of interest and characterize it.
Can gene tools provide Morpholino oligos?
Gene Tools can provide Morpholino oligos with modifications on the 3′ and/or 5′ ends of the oligos. Available structures are shown below. The following filters and dichroic mirror from Chroma Technology Corp. match the excitation and emission of Gene Tools Blue.
Who makes Morpholino antisense oligos?
Gene Tools makes Morpholino antisense oligos. Morpholino oligos bind to complementary RNA and get in the way of processes; they can knock down gene expression, modify RNA splicing or inhibit miRNA activity and maturation. Morpholinos are the premier knockdown tools used in developmental biology labs,…
What is a Morpholino standard control oligo?
Standard Control: A Morpholino Standard Control oligo is offered at a reduced price [$100/100 nmol] to minimize the expense of antisense experiments. This Standard Control oligo is a 25-mer having the sequence:
What is a Morpholino?
Morpholinos are the premier knockdown tools used in developmental biology labs, the best RNA-blocking reagents for cells in culture and, as Vivo-Morpholinos, the most specific delivery-enhanced oligos available for other animal models. We are the sole commercial manufacturer selling research quantities of Morpholinos world-wide.