Table of Contents

Custom Morpholinos

Morpholino Antisense Oligos function by binding to pre-messenger or messenger RNA, blocking access to the RNA. Morpholinos function solely by a steric block mechanism, so they do not require or use RNase H for their activity. When properly selected and targeted, Morpholinos generally afford equal or better efficacy and greatly superior specificity than any other antisense structural type. However, inappropriately selected or targeted Morpholinos will have little or no activity. Accordingly, Gene Tools provides an Oligo Design Service at no added cost. Gene Tools follows our Targeting Guidlines to select appropriate oligo sequences suitable for inhibiting the activity of the genes of your choice.

For quantity and price information on custom-sequence Morpholino antisense oligos go to Prices.

For selecting and ordering Morpholino oligos visit our Online Store.

---

Control Oligos

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:


This oligo should have no target and no significant biological activity - except in reticulocytes from thallasemic humans having a splice-generating mutation at position 705 in beta-globin pre-mRNA. In those cells this oligo will correct a splicing error and thereby generate a correctly-spliced mRNA which codes for normal beta-globin chains. For further information on this splice-correction process go to: Positive Antisense Test System.

For quantity and price information on this Standard Control oligo go to Prices.

Custom Controls: Custom-sequence Morpholino control oligos can also be ordered. If the standard control oligo is unsuitable for your experiments, we recommend using the invert of your antisense sequence as your negative control. This invert-antisense has the advantage of having the same length and base composition as your antisense oligo.

Here is an example based on an arbitrary antisense sequence:

antisense sequence: 5'-AAC GAA CGA ACG AAC GAA CGA ACG C

invert antisense: 5'-CGC AAG CAA GCA AGC AAG CAA GCA A

A less desirable negative control is the sense sequence. Sometimes odd expression modulation is observed with sense controls, such as upregulation of the gene of interest (hypothetically due to DNA interactions).

antisense sequence: 5'-AAC GAA CGA ACG AAC GAA CGA ACG C

sense sequence: 5'-GCG TTC GTT CGT TCG TTC GTT CGT T

For rigorous specificity studies we recommend using a mis-pair control oligo, with a sequence nearly the same as the antisense sequence but incorporating five mis-pairs along the length of the oligo. Experiments at Gene Tools have shown that the best five-mispair controls are those in which three out of the five misparings are Cs and/or Gs, as illustrated below:

antisense sequence: 5'-AAC GAA CGA ACG AAC GAA CGA ACG C

5-mis-pair control: 5'-AAg GAA gGA ACc AAC GAA CcA AgG C

These mis-paired control oligos provide a much more stringent and realistic assessment of the true sequence specificity of your antisense oligo. It is noteworthy that in studies of sequence specificity utilizing such mis-paired controls, Morpholino oligos were found to afford high-specificity target inhibition over a concentration range several hundred fold greater than that afforded by corresponding Phosphorothioate oligos (Summerton et al., 1997; Summerton & Weller, 1997a).

For quantity and price information on custom-sequence Morpholino control oligos go to Prices.

---

End Modifications

Gene Tools can provide Morpholino oligos with modifications on the 3' and/or 5' ends of the oligos. Available structures are shown in the chart below.

• LISSAMINE NOTE: Lissamine (sulforhodamine B) can decrease the aqueous solubility of many Morpholino sequences. If a green fluor will suffice, we suggest you have your oligos labelled with carboxyfluorescein.


• BIOTIN NOTE: we do not have a protocol for using the biotin modified Morpholinos as in situ probes. Doing so will require development of a protocol to bind the biotinylated Morpholino to complementary mRNA, then wash away unbound Morpholino without washing away the Morpholino & mRNA heteroduplex. So far we do not know of a technique that will do this.

---