Charged PNA Monomers for Abasic Probe Stability
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Solution Overview
Problem
Peptide nucleic acid (PNA) probes with 'blank' or 'abasic' units, when attached to solid supports, tend to deform and lose specificity and sensitivity due to the absence of a nucleobase, leading to non-specific binding and variable detection efficiency in genetic analysis methods.
Innovation Solution
Incorporating a charged moiety or a moiety capable of carrying a charge at a predetermined pH into PNA monomers to enhance the stability and conformational integrity of PNA molecules, thereby improving their specificity and sensitivity when used in genetic analysis and nucleobase characterization assays.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If a PNA probe contains a blank or abasic unit, then the probe can be used for characterisation of nucleobases, but the probe deforms and loses specificity and sensitivity when attached to solid supports
Solution Approach 1:
The patent introduces a charged moiety at the gamma position of the PNA monomer, changing the chemical parameter of the backbone. This modification stabilizes the PNA probe structure when attached to solid supports, preventing deformation while maintaining the blank unit's characterisation function, thus resolving the contradiction between versatility and reliability
Solution Approach 2:
The patent creates a composite PNA structure by combining the peptide backbone with a charged moiety (such as carboxylic acid, phosphonic acid, or phosphoric acid groups) at the gamma position. This composite structure provides both the characterisation capability of the blank unit and the structural stability provided by the charged moiety, eliminating probe deformation
2Adaptability or versatility
If a PNA probe contains a blank or abasic unit, then the probe can be used for genetic analysis, but the probe binds non-specifically to nucleobases
Solution Approach 1:
By modifying the PNA monomer with a charged moiety at the gamma position, the patent changes the electrostatic and structural parameters of the probe. This modification enhances the probe's specificity for complementary nucleobases while reducing non-specific binding, allowing the blank unit to maintain its genetic analysis function without causing harmful non-specific interactions
3Adaptability or versatility
If a PNA probe contains a blank or abasic unit, then the probe can be used for sequencing applications, but the detection efficiency becomes variable
Solution Approach 1:
The patent modifies the PNA probe by introducing a charged moiety at the gamma position, which stabilizes the probe's hybridization to the target sequence. This parameter change ensures consistent and reliable detection efficiency throughout the sequencing process, eliminating the variability associated with blank units while maintaining sequencing capability
Data Source
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AI summary
Disclosed herein are improved PNA based monomers, nucleobases, oligomers and probes for use in a variety of different methods of analysing nucleic acids. Further, the disclosure provides methods of preparing the modified and improved PNA molecules as well as methods of using the same.