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Antisense Oligos

Antisense Oligos
 
 
In antisense technology short synthetic oligonucleotides are supposed to hybridize to a certain sequence of the mRNA (drug target) thereby interfering with the mRNA processing. In order to equip an oligonucleotide with drug-like properties, chemical modification is inevitable. Microsynth provides most prominent modification strategies that are used in antisense technology like:
  • 2'-O-Methyl
  • 2'-Fluoro
  • MOE (2'-methoxyethyl)
  • LNA (locked nucleic acid)
  • S-cEt (constrained ethyl, on request)
  • PTO (phosphorothioate)
  • Gapmers
  • GalNAc
 

Features and Benefits

 
 
High Quality
  • All ASOs are MALDI-TOF MS controlled before shipment
  • Renowned ASO supplier (primarily for drug discovery phase) to top research institutes and pharma/biotech companies

 

Fast Production
  • Synthesis scales up to 1.0 µmol including HPLC are shipped within 3-5 days
 
Excellent Technical Support
  • Trained and experienced scientists are happy to support you
 
 
 
Cost Effective
  • Competitive pricing
  • High guaranteed yields

 

Convenient
  • Analytical HPLC available
  • Certificate of Analysis available
 
User-friendly Online Ordering System
  • Easy-to-use online portal with a series of helpful tools (e.g. order tracking & history, convenient search and re-order option)
 

Overview

 
Microsynth can offer you the following modifications:
 

 

  • PTO (phosphorothioates) modifications: PTOs contain one sulfur atom in place of an oxygen atom in the internucleotide linkage of DNA or RNA. This modification of the normal phosphodiester backbone is characterized by an increased cell uptake, high nuclease resistance and elicitation of RNAse H activity.
  • 2'-O-Me-RNA modifications: The incorporation of 2'-O-Methyl RNA nucleotides induces a resistance to a wide variety of nucleases, in particular RNase. Furthermore, 2’-OMe oligonucleotides show slightly increased affinity towards their complementary mRNA target sequence, thereby forming more stable hybrid duplexes compared to their non-modified DNA or RNA counterparts. This enables the formation of more stable hybrids with complementary RNA strands than would be the case for non-modified DNA and RNA sequences.
  • 2'-MOE-RNA modifications: Oligonucleotides incorporating 2'-O-methoxyethyl (MOE)-modified nucleotides, can support most, if not all antisense mechanisms of action. Further key distinctive characteristics are nuclease resistance, lower toxicity, superior target binding specificity, as well as increased affinity towards complementary RNA. For more detailed information about 2'-MOE antisense oligonucleotides from Microsynth, please see the flyer on the right side.
  • LNA modifications: LNA containing oligonucleotides offer substantially increased affinity for its complementary strand, compared to traditional DNA or RNA oligonucleotides. This and the concomitant high nuclease resistance of LNAs results in unprecedented sensitivity and specificity and makes LNA™ oligonucleotides ideal for use in antisense applications.

  • S-cEt modifications: cEt oligonucleotides (constrained ethyl nucleotides) are an advancement of LNA oligonucleotides. They have been developed for antisense applications and are mainly used in gapmers. Such ASOs show superior stability towards nuclease degradation compared to LNA without compromising binding selectivity or hybridization stability.
  • N-acetylgalactosamine (GalNAc): Conjugation of N-acetylgalactosamine (GalNAc) has become a major clinical strategy for delivery of oligonucleotides to hepatocytes (liver cells). Such conjugates are efficiently internalized by binding to the asialoglycoprotein receptor (ASGR), which exhibits high affinity for N-acetyl galactosamine terminated oligosaccharides.

 

To learn more about specifications of these five types of modifications, please see the table below.


Key Specifications:

Modifications

Position

Synthesis scale [µmol] Purification1

5’

3’

Int.

0.04

0.2

1

15

HPLC + Dialysis

PTO

   

x

x

x

x

x

x

2’-O-Me-RNA

x

x

x

x

x

x

x

x

2'-MOE-RNA x x x x x x x x
LNA x x x x x x x x
GalNAc   x     x x x x
 
 

1 We strongly recommend to order ASOs with “HPLC + Dialysis” purification. The Na+ salt exchange is necessary to remove toxic salts from HPLC purification.

Synthesis Scales

Full 2' OMe, 2' MOE in combination with or without PTO

Desalted

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 3 15 2-3
0.2 µmol 10 50 2-3
1.0 µmol 30 150 2-3
15 µmol 600 3'000 3-5
 

HPLC

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 1 5 3-4
0.2 µmol 3 15 3-4
1.0 µmol 10 50 3-4
15 µmol 300 1'500 4-6
 

HPLC & Dialysis

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 - - -
0.2 µmol 3 15 3-5
1.0 µmol 10 50 3-5
15 µmol 250 1'250 5-7
 

PAGE

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 0.5 2.5 3-4
0.2 µmol 2 10 3-4
1.0 µmol 7 35 3-4
15 µmol - - -
 

Gapmers (including DNA and 2' OMe or 2' MOE)

HPLC

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 1 5 3-4
0.2 µmol 3 15 3-4
1.0 µmol 8 40 3-4
15 µmol 300 1'500 4-6
 

HPLC & Dialysis

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 - - -
0.2 µmol 3 15 3-5
1.0 µmol 8 40 3-5
15 µmol 250 1'250 5-7
 

PAGE

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 0.5 2.5 3-4
0.2 µmol 2 10 3-4
1.0 µmol 7 35 3-4
15 µmol - - -
 

Gapmers (including DNA and LNA)

HPLC

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 1 5 3-4
0.2 µmol 2 10 3-4
1.0 µmol 6 30 3-4
15 µmol 250 1'250 4-6
 

HPLC & Dialysis

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 - - -
0.2 µmol 2 10 3-5
1.0 µmol 6 30 3-5
15 µmol 200 1'000 5-7
 

PAGE

Synthesis scale1 Length Restriction Guaranteed Yield2 Production Time [wd]
[OD260] [nmol]3
0.04 µmol 13 - 30 0.5 2.5 3-4
0.2 µmol 2 10 3-4
1.0 µmol 5 25 3-4
15 µmol - - -
 
1 The synthesis scale represents the initial amount of 3' bases (starting material).
2 Our guaranteed and average yields are measured in OD.
3 Yields indicated in nmol represent an example calculation for a 20mer. For this calculation the following rule of thumb equation was applied: nmol of oligo = OD x 100/length of oligo. Please note that this calculation is based on sequences with virtually homogenous distribution of the 4 DNA bases; it may vary for sequences with high GC contents >70% etc.

How to Order

 
How to Order?
  • Enter our webshop
  • Click on DNA in the blue "DNA/RNA Synthesis" domain
  • Select either Normal Entry in order to type or copy/paste the desired sequence information etc. or alternatively select Upload Entry by using our convenient Excel template (can be downloaded from our webshop).
 
PTO modified oligonucleotides: please mark PTO bonds by inserting an asterisk between the bases, e.g. AAA*T*G*CCC*AAA.
 
 
2’-O-Me-RNA (Selective incorporation of 2’-O-Me-RNA bases into DNA):
  • Enter 5,6,7,8 for each distinct base to be replaced inside the sequence
  • Define Inner Modification (5=…) by selecting the desired modification from the drop down menu (e.g. 5=…2’-O-Methyl-RNA-A, 6=…2’-O-Methyl-RNA-C etc.)
 
2’-MOE-RNA (Selective incorporation of 2’-MOE-RNA bases into DNA):
  • Enter 5,6,7,8 for each distinct base to be replaced inside the sequence
  • Define Inner Modification (5=…) by selecting the desired modification from the drop down menu (e.g. 5=…2’-Methoxyethyl-ribo-A, 6=…2’-Methoxyethyl-ribo-mC etc.)
 
LNA (Selective incorporation of LNA bases into DNA):
  • Enter 5,6,7,8 for each distinct base to be replaced inside the sequence
  • Define Inner Modification (5=…) by selecting the desired modification from the drop down menu (e.g. 5=…LNA-A, 6=…LNA-C etc.)
 
S-cEt
  • cEt ASO are available on request only. Ordering instructions follow together with the offer.