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Artificial Base Pair Systems for Expanded Genetic Alphabets

Non-natural nucleobase systems for expanded information storage, PCR and diagnostics, aptamer evolution, site-specific labeling, protein engineering, and synthetic biology beyond A, T(U), G and C.

Benner dP:dZ Benner dB:dS dIsoC:dIsoG dNaM:dTPT3 dDs:dPx Hachimoji DNA

Engineering New Pairing Rules Beyond Nature’s Four Bases

Artificial base pair systems introduce non-natural nucleobases that recognize complementary partners while remaining embedded in a DNA- or RNA-like scaffold. Unlike backbone or sugar modifications, these systems change the information-bearing bases and create additional pairing relationships beyond A:T(U) and G:C.

Bio-Synthesis supports project review for Benner AEGIS dP:dZ and dB:dS, dIsoC:dIsoG, Hirao dDs:dPx, Romesberg dNaM:dTPT3, Hachimoji systems and selected emerging bases.

Artificial base pair structures showing hydrogen-bonded and hydrophobic unnatural base pairs

Additional Information

Adds orthogonal genetic letters.

Selective Recognition

Hydrogen-bonded or hydrophobic pairing.

Enzymatic Processing

Selected systems support replication or transcription.

Functional Expansion

Enables aptamers, labeling and protein engineering.

Nomenclature: dP, dZ, dB, dS, dIsoC, dIsoG, dDs, dPx, dNaM and dTPT3.

Available Artificial Nucleobases

Artificial Base Complement Chemistry Family DNA Oligo RNA Internal 5′ 3′ Notes
dP dZ Benner AEGIS Research Confirm monomer and scale.
dZ dP Benner AEGIS Research Protecting-group and MS review.
dB dS Benner AEGIS Research Sequence context may affect handling.
dS dB Benner AEGIS Research Confirm amidite and deprotection route.
dIsoC dIsoG Iso Base Pair Research Useful for orthogonal controls.
dIsoG dIsoC Iso Base Pair Research Tautomerism should be considered.
dDs dPx Hirao Research Enzymatic Specialty monomer and purification review.
dPx dDs Hirao Research Research Confirm pair generation and substrate form.
dNaM dTPT3 Romesberg Research Enzymatic Hydrophobic base; analytics may differ.
dTPT3 dNaM Romesberg Research Research Confirm monomer source and project scale.

Notes: Check marks indicate feasible placement formats for project review, not blanket stock availability. RNA compatibility may require enzymatic workflows or dedicated ribonucleoside chemistry. Additional research bases such as dPn, dPa, d5SICS, dMMO2, s, y, ImN, NaO and CTPT3 can be evaluated when a structure or publication is provided.

Choose a Base Pair System by Research Objective

Select your project goal to see the recommended artificial base-pair family, why it fits, common alternatives, practical applications and the main technical consideration.

Choose a research objective below to update the recommendation.
Primary Recommendation

Benner dP:dZ

dP:dZ is the strongest general starting point for expanding genetic information while preserving a Watson–Crick-like hydrogen-bonding framework. It is central to AEGIS and also contributes to larger expanded-alphabet systems.

Benner dP:dZ dB:dS Hachimoji
Information Storage ★★★★★
PCR History ★★★★★
Diagnostics ★★★★★
Alternative Systems
System Best Use
dB:dS Orthogonal primer and assembly workflows
Hachimoji Eight-letter information systems
Typical Applications
  • Expanded DNA
  • Information storage
  • PCR and sequencing research
  • Orthogonal molecular systems
Main Consideration

Replication fidelity and sequence-context performance must be validated in the complete enzyme system.

Primary Recommendation

Benner dP:dZ or dIsoC:dIsoG

These hydrogen-bonded systems are strong starting points for orthogonal amplification, internal controls and diagnostic workflows. dP:dZ offers broader expanded-alphabet potential, while dIsoC:dIsoG is especially useful for control sequences and detection systems.

dP:dZ dIsoC:dIsoG dB:dS

PCR

★★★★★

Diagnostics

★★★★★

Orthogonality

★★★★☆
Alternative Systems
System Best Use
dB:dS Orthogonal primers and gene assembly
dDs:dPx Optimized expanded-alphabet PCR
Typical Applications
  • Orthogonal primers
  • Diagnostic controls
  • Multiplex assays
  • Expanded templates
Main Consideration

“PCR compatible” is polymerase-, sequence- and cycle-dependent.

Primary Recommendation

Hirao dDs:dPx

dDs:dPx is the strongest starting point for expanded-alphabet aptamer discovery because the hydrophobic base pair adds chemical diversity and has a strong history in ExSELEX and high-affinity aptamer generation.

dDs:dPx dNaM:dTPT3 dP:dZ

Aptamers

★★★★★

Chemical Diversity

★★★★★

PCR

★★★★☆
Alternative Systems
System Best Use
dNaM:dTPT3 Hydrophobic diversity and expanded biology
dP:dZ Hydrogen-bonded expanded libraries
Typical Applications
  • ExSELEX
  • High-affinity binders
  • Protein and small-molecule targets
  • Functionalized libraries
Main Consideration

Selection value depends on reliable amplification and retention of the artificial pair.

Primary Recommendation

Romesberg dNaM:dTPT3

dNaM:dTPT3 has the strongest history in expanded codons, semi-synthetic organisms and noncanonical amino acid incorporation. It is the preferred starting point when the goal extends from DNA storage into transcription and translation.

dNaM:dTPT3 Expanded Codons Novel Proteins

Protein Engineering

★★★★★

In Vivo Use

★★★★★

System Complexity

Very High
Alternative Systems
System Best Use
dDs:dPx Functional nucleic acid labeling
Hachimoji Expanded information research
Typical Applications
  • Expanded codons
  • Noncanonical amino acids
  • Novel protein expression
  • Translation-system engineering
Main Consideration

The entire replication, transcription and translation system must be engineered together.

Primary Recommendation

Hirao dDs:dPx

Hirao-family systems are well suited to site-specific labeling because functionalized artificial-base substrates can be incorporated at defined positions through engineered replication or transcription workflows.

dDs:dPx dDs:dPa Functional Triphosphates

Labeling

★★★★★

RNA Production

★★★★☆

Functional Diversity

★★★★★
Alternative Systems
System Best Use
dNaM:dTPT3 Expanded transcription systems
dP:dZ Orthogonal template design
Typical Applications
  • Site-specific dyes
  • Biotinylated RNA or DNA
  • Functional nucleic acids
  • Imaging and binding studies
Main Consideration

Chemical incorporation and enzyme-mediated labeling require different substrates and workflows.

Primary Recommendation

Romesberg dNaM:dTPT3

dNaM:dTPT3 is the most established system for maintaining an unnatural base pair in living cells and retrieving expanded information through transcription and translation.

dNaM:dTPT3 Semi-Synthetic Organism Expanded Translation

In Vivo Retention

★★★★★

Translation

★★★★★

Complexity

Very High
Alternative Systems
System Best Use
Hachimoji Expanded information systems
dP:dZ Hydrogen-bonded synthetic genetics
Typical Applications
  • In vivo pair retention
  • Expanded transcription
  • Novel codons
  • Noncanonical proteins
Main Consideration

Triphosphate transport, replication fidelity and cellular fitness all influence success.

Explore Major Expanded-Alphabet Systems

Select any tab below to open its artificial base pair profile.
P:Z

Benner AEGIS dP:dZ

dP and dZ form an orthogonal hydrogen-bonded pair with Watson–Crick-like geometry.

Individual Bases

dP, dZ

Pairing

Hydrogen Bonds

Best Known For

AEGIS

View dP:dZ Guide →

Applications

  • Six-letter DNA
  • PCR and sequencing
  • Aptamers
  • Diagnostics

Order as

  • dP-containing oligo
  • dZ-containing oligo
  • Matched duplex

Advantages

  • Orthogonal pairing
  • Natural-like geometry

Use Caution

  • Sequence context
  • Polymerase specificity
B:S

Benner AEGIS dB:dS

dB and dS form a second AEGIS pair used in orthogonal primer and assembly workflows.

Individual Bases

dB, dS

Pairing

Hydrogen Bonds

Best Known For

Orthogonal Primers

View dB:dS Guide →

Applications

  • Gene assembly
  • Orthogonal primers
  • Transliteration

Order as

  • dB-containing oligo
  • dS-containing oligo

Advantages

  • Reduced cross-talk
  • Expanded sequence space

Use Caution

  • Less common than P:Z
  • Enzyme-specific
iC:iG

dIsoC:dIsoG

A historically important hydrogen-bonded expanded pair used in diagnostics and PCR research.

Bases

dIsoC, dIsoG

Pairing

Hydrogen Bonds

Best Known For

Diagnostics

View IsoC:IsoG Guide →

Applications

  • Diagnostic controls
  • Orthogonal amplification

Order as

  • IsoG tautomerism
  • Sequence context
NaM

dNaM:dTPT3

A hydrophobic pair associated with semi-synthetic organisms and expanded translation.

Bases

dNaM, dTPT3

Pairing

Hydrophobic

Best Known For

Expanded Codons

View NaM:TPT3 Guide →

Applications

  • Semi-synthetic organisms
  • Protein engineering

Order as

  • Requires engineered system
  • Triphosphate transport
Ds:Px

dDs:dPx

A Hirao hydrophobic pair used in PCR, ExSELEX, high-affinity aptamers and labeling.

Bases

dDs, dPx

Pairing

Hydrophobic

Best Known For

ExSELEX

View Ds:Px Guide →

Applications

  • Aptamers
  • Site-specific labeling
  • PCR

Order as

  • dPn
  • dPa
  • s and y
8

Hachimoji DNA and RNA

Eight genetic letters forming four orthogonal pairs for predictable expanded information systems.

Alphabet

Eight Letters

Pairs

Four

Best Known For

Expanded Genetics

View Hachimoji Guide →

Applications

  • Information storage
  • Structural biology

Use Caution

  • Multiple artificial bases
  • Complex enzyme system
HB

Hydrophobic Base Pair Systems

Recognition based on shape, packing and exclusion of water.

Examples

  • dNaM:dTPT3
  • dDs:dPx
  • d5SICS and dMMO2 families

Selection

  • Aptamers: Ds:Px
  • Translation: NaM:TPT3
PCR

PCR-Compatible Systems

Selected artificial pairs can be retained under optimized amplification conditions.

Candidates

  • dP:dZ
  • dB:dS
  • dIsoC:dIsoG
  • dDs:dPx
  • dNaM:dTPT3

Controls

  • Pair-retention assay
  • Polymerase comparison
  • Sequence verification
DX

Diagnostic Systems

Artificial pairs create orthogonal controls, primers and probes with reduced natural-genome cross-talk.

Common Choices

  • dP:dZ
  • dB:dS
  • dIsoC:dIsoG

Applications

  • Internal controls
  • Multiplex workflows
  • Capture probes
NEW

Emerging Artificial Bases

New C-nucleosides, hydrolysis-stable partners and expanded aromatic systems continue to emerge.

Examples

  • CTPT3-related systems
  • ImN:NaO
  • xDNA and yDNA

Project Review

  • Provide structure
  • Provide publication
  • Define scale and QC

From Artificial Base Selection to Final Application

A successful artificial-base project usually combines chemical synthesis with assay-specific enzymology and analytical confirmation.

Artificial Base → Synthesis → PCR or Transcription → Application

The workflow can be adapted for direct chemical incorporation, enzyme-mediated labeling, aptamer selection, diagnostics or expanded-translation studies.

1

Select the Pair

Choose dP:dZ, dB:dS, IsoC:IsoG, Ds:Px, NaM:TPT3 or another system based on the intended application.

2

Synthesize the Oligonucleotide

Define sequence, artificial-base position, scale, purification, terminal labels and analytical requirements.

3

Amplify or Transcribe

Use a compatible polymerase or transcription system and verify retention of the artificial base pair.

4

Apply & Validate

Use the product in diagnostics, aptamer selection, labeling, protein engineering or expanded-genetic studies.

Critical control: confirm artificial-base identity and retention at each stage. Chemical incorporation, PCR fidelity, transcription efficiency and biological function are separate validation steps.

Artificial Base Pair Family Comparison

Relative ratings summarize the maturity and typical usefulness of each family for common applications. Actual performance remains polymerase-, sequence- and workflow-dependent.

System Pairing Mechanism Polymerase / PCR Aptamers Protein Engineering Diagnostics Semi-Synthetic Organisms Primary Strength
Benner AEGIS dP:dZ Hydrogen bonding ★★★★★ ★★★★★ ★★★★★ ★★★★★ ★★★★★ Expanded information, PCR and diagnostics
Benner AEGIS dB:dS Hydrogen bonding ★★★★ ★★★★★ ★★★★★ ★★★★ ★★★★★ Orthogonal primers and gene assembly
dIsoC:dIsoG Hydrogen bonding ★★★★ ★★★★★ ★★★★★ ★★★★★ ★★★★ Orthogonal controls and diagnostics
Hirao dDs:dPx Hydrophobic packing ★★★★ ★★★★★ ★★★★ ★★★★★ ★★★★★ Aptamer evolution and labeling
Romesberg dNaM:dTPT3 Hydrophobic packing ★★★★★ ★★★★★ ★★★★★ ★★★★ ★★★★★ Expanded translation and in vivo systems

Ratings are directional, not absolute specifications. Polymerase choice, sequence context, cycle count, substrate form and assay design can materially change performance.

Representative Applications

Artificial base pairs expand the chemical and informational capabilities of nucleic acids across diagnostics, molecular evolution, synthetic biology and protein engineering.

DNA

Expanded Information Storage

Add orthogonal genetic letters and pairing relationships beyond natural DNA.

Common systems: AEGIS, Hachimoji
PCR

PCR & Diagnostics

Create orthogonal controls, primers and detection systems with reduced natural cross-talk.

Common systems: AEGIS, IsoC:IsoG
APT

Aptamer Evolution

Expand library chemistry with hydrophobic and orthogonal nucleobases.

Common systems: Ds:Px, NaM:TPT3, P:Z
PRO

Protein Engineering

Create expanded codons for noncanonical amino acid incorporation.

Common system: NaM:TPT3
TAG

Site-Specific Labeling

Direct functional-group incorporation into DNA or RNA at defined positions.

Common systems: Hirao pairs
CELL

Semi-Synthetic Organisms

Retain, transcribe and retrieve expanded genetic information in living cells.

Common system: NaM:TPT3

Combine Artificial Bases with Functional Modifications

Fluorophores

Labeled probes and transcripts.

Click Handles

Modular ligands and surfaces.

Biotin

Capture and enrichment.

Backbone Chemistries

PS, LNA/BNA or other compatible modifications.

FAQ

What are the Benner AEGIS bases?
 dP, dZ, dB and dS, forming dP:dZ and dB:dS.
How do hydrophobic pairs differ?
 They rely more on shape and packing than classical hydrogen bonds.
What is Hachimoji DNA?
An eight-letter genetic system containing four orthogonal pairs.
Can artificial bases be used in RNA?
 Some systems support enzymatic transcription; chemical RNA synthesis requires separate review.
Can artificial bases carry dyes or click handles?
 Yes, by chemical or enzyme-mediated workflows depending on the system.
How should bases be specified?
 Use exact notation such as dP, dZ, dB, dS, dIsoC, dIsoG, dDs, dPx, dNaM and dTPT3.
Can artificial bases be amplified by PCR?
 Several systems can, under optimized and system-specific conditions.
Does Bio-Synthesis guarantee polymerase performance?
 No. Polymerase performance must be validated in the complete customer assay.

Need help planning an artificial-base oligonucleotide?

Send the sequence, exact artificial-base names, pair family, incorporation positions, workflow, scale, purification target, labeling needs and analytical expectations.

What to Send

  • Sequence and notation
  • Pairing partner
  • Chemical or enzymatic workflow
  • Scale, purification and QC

What We Review

Monomer availability, incorporation chemistry, purification, MS compatibility and duplex requirements.

Quality Systems & Custom Project Support

QMS

ISO-Supported Advanced Oligonucleotide Manufacturing

Controlled production, project-specific purification, analytical QC, documentation and packaging.

ISO 9001:2015 Quality management
ISO 13485:2016 Medical-device framework
ISO 14001 Environmental management
Analytical QC HPLC/UPLC, MS where compatible, OD and COA

Selected Literature by Research Platform

Benner AEGIS & Hachimoji

  • Alternative Watson–Crick systems and dP:dZ structural studies.
  • Hachimoji DNA and RNA.
  • Expanded-alphabet sequencing and gene assembly.

Hirao

  • Ds-based pairs for PCR, labeling and ExSELEX.

Romesberg

  • NaM:TPT3 in semi-synthetic organisms and expanded translation.

Historical & Emerging

  • IsoC:IsoG, CTPT3, ImN:NaO, xDNA and yDNA.

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