Custom Circular RNA Synthesis

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Custom Circular RNA Synthesis

For protein coding and non-coding RNA

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Custom Circular RNA (circRNA) Service- Scarless circRNA and circRNA with Scar

Unlock groundbreaking advancements in research, therapeutics, and diagnostics with GenScript's cutting-edge custom circular RNA (circRNA) synthesis services. Designed to drive innovation, our state-of-the-art circRNA solutions—featuring circRNA with Scar by PIE, Scarless circRNA by PIE, and Scarless circRNA by T4 ligation—deliver unmatched precision, stability, and optimized expression.

Our versatile circRNA platform caters to both protein-coding (circRNA with Scar by PIE, Scarless circRNA by PIE) and non-coding circRNAs (Scarless circRNA by T4 ligation), enabling transformative applications in gene editing, cellular process control, immune modulation, gene therapy, vaccine development, and biomarker discovery.

Empower your next breakthrough with GenScript’s engineered circRNA solutions, tailored to meet the demands of modern science and medicine. Elevate your research and therapeutic potential with precision-engineered circRNA designed for innovation and impact.

Rush mRNA Synthesis New!

GMP-like mRNA Manufacturing Services

Catalog IVT mRNA, circRNA, saRNA, LNP products

mRNA QC Analysis

GenScript Cap Technology

Circular RNA Synthesis

Self-Amplifying RNA

LNP Formulation

Targeted LNP Formulation

ReadyEdit LNP Formulation

Benefits of circRNA synthesis with GenScript

GenScript is the trusted choice for scientists worldwide, offering flexible, high-quality circular RNA synthesis solutions. Our proprietary circRNA vectors enables high circularization efficiency and purity.with customizable designs support to support your research needs. Scalable from 100 μg to 15 mg, ≥ 80% HPLC purified delivered in three weeks. We ensure precision , reliability, and high-performance RNA. The result? Enhanced RNA stability, reduced immunogenicity, prolonged protein expression, and superior translation efficiency, driving innovation in research and therapeutics.

Customizable designs - For protein coding and non-coding applications.

Scales ranging from 100 μg to 15 mg to suit projects of all sizes.

Cell-specific IRES options optimized for Specific tissue/cell-lines (CVB3, HRV-B3, EV-B107, CVB1 or customer designed IRES)

Delivery of circRNA (≥ 80%, HPLC-purified) in 3 weeks

Select the method best suited for your target application

Enabling Precision and Efficiency – Scarless circRNA and circRNA with Scar

At GenScript, we provide three circRNA custom synthesis methods, ensuring the precise solution your research demands. Each method offers specific advantages tailored to different applications.

circRNA with Scar by PIE- Self circularization

Scarless circRNA by T4-Ligation

Application

For protein translation in coding applications

For protein translation in coding applications w/ improved stability and protein expression vs. circRNA with scar

For noncoding RNA applications

Description

Generates circRNA with a small scar sequence

Generates circRNA with no scar sequence

Uses T4 ligase for seamless circularization

Total Sequence Size

7 kb

5 kb

2 kb

Final circRNA sequence

IRES+Exon+Linker (Scar) Sequence+ORF

IRES+ORF

Customer provided sequence

Custom circRNA Synthesis Workflow: From Design to Delivery

Design selection

Protein-coding application:

circRNA with Scar by PIE

Scarless circRNA by PIE

Non-coding application:

Scarless circRNA by T4 Ligation

Customization

Non-modified or 5% m6A

IRES options-CVB3, HRVB3, EV-B107, CVB1, customer-designed IRES

Sequence length 7 Kb

Scale: 0.1-15 mg

QC tests

Purity (HPLC), ID (gel electrophoresis), impurity & safety checks (endotoxin assay)

Research-grade Standard QC

RUO upgrade

Custom QC tests

Delivery

Receive high-purity (≥ 80%, HPLC-purified), ready-to-use circRNA

QC Section

Identification

Appearance

Visual inspection

Clear and free of foreign particles

RNA length

Agarose gel electrophoresis

Expected size band detected

RNaseR degradation test

Not be degraded by RNase R

RNA content

UV absorbance

Target ± 5%

pH paper

Target ± 0.5

Buffer specification

Client spec

N/A

Purity

A 260/280 ratio

UV Spec

1.70 ~ 2.30

Purity by HPLC

HPLC

≥80%

Impurity

Total protein residue

NanoOrange assay

≤ 1%

Plasmid DNA residue

qPCR

≤ 0.05%

Safety

Endotoxin

Semiquantitative

< 10 EU/mg

Quantitative

Bioburden

Direct inoculation

No growth after 48 hrs

Case Study

The translation efficiency and stability of different RNA formats (Linear mRNA, circ-Scar, and circ-Scarless) in two cell lines: HEK293T (human embryonic kidney cells) and Jurkat cells (human T lymphocyte cells). Superior expression compared to circ-Scar. Expression peaks at 12 hours and remains high, with gradual decline after 6 days, but still outperforms both Circ-Scar and linear mRNA.

RNA Copy Number by RT-PCR

RNA Expression by Fluorescence Intensity

Circular eGFP- RNA (nonmodified) and Linear eGFP-mRNA (100% N1-me-pseU modified) were transfected with equal molar quantity in A549 cells. Coding RNA abundance was measured by qPCR, and GFP intensity was measured by flow cytometry. Results demonstrated that circRNA has higher stability, slower degradation, compared to linear IVT mRNA. circRNA also exhibits higher protein expression level and duration compared to linear IVT mRNA

GenScript circRNA demonstrates lower immunogenicity than mRNA. For the IL-6 assay, the ELISA method was performed using the Abcam Human IL-6 ELISA Kit (ab46027). For IFN-α and NF-κB assays, A549 Dual cells (InvivoGen) were transfected with test mRNA. IFN-α expression was measured by relative luminescence for ISG54 activity following the QUANTI-Luc™/Gaussia kit instructions. NF-κB levels were measured by optical density at 630 nm from the culture medium using the QUANTI-Blue™ kit.

GenScript circRNA shows higher initial expression and cumulative expression than both mRNA and circular RNA prepared with a published method. A549 cells were transfected with circRNAs and mRNA encoding eGFP. circRNAs were prepared using GenScript’s proprietary method and using methods published in Nat Commun 9, 2629 (2018). Fluorescence was analyzed via flow cytometry for up to 6 days following transfection (10,000 cells were analyzed per condition at each time point). A549 cell doubling time = 22hr.

EGFP circRNA expression in A549 cells, 24 hrs

EGFP circRNA expression in HEK293 cells, 24 hrs

GenScript HPLC-purified circRNA shows higher expression than both crude circRNA and circRNA prepared with a published method. Cells were transfected with circRNAs encoding eGFP prepared using GenScript’s proprietary method and using methods published in Nat Commun 9, 2629 (2018). Fluorescence was analyzed via flow cytometry.

Popular off-the-shelf circRNA catalogs available now!

Try our catalog mRNA expressing eGFP or F-Luciferase, delivered in just 5 days!

Resources

Circular RNA Case Study Report

Circular RNA introduction, GenScript offering details, and case studies showing increased stability, prolonged expression, and lower immunogenicity compared to linear IVT mRNA.

Circular RNA Beginner’s Guide_Infographic

Learn about key structural components of circRNA, the process by which circRNA is translated into protein, and more.

FAQ-Circular RNA Synthesis

circRNA with Scar by PIE is suited for protein translation for coding applications where a small scar sequence is acceptable.

Scarless PIE offers improved stability and efficiency for protein-coding RNA vs. circRNA with scar.

T4 ligation is perfect for fully scarless RNA. Preferable for non-coding applications.

Scarless circRNA eliminates unwanted sequence scars, resulting in a more natural RNA structure with improved stability and efficiency. circRNA with Scar includes strong IRES activity and effective expression in various models.

GenScript’s circRNA synthesis process features proprietary circularization techniques and HPLC purification, ensuring the highest purity and RNA integrity.

Absolutely! We offer cell-specific IRES options optimized for various applications, including gene therapy and functional studies.

circRNA offers enhanced stability, prolonged protein expression, lower immunogenicity, and reduced degradation, making it ideal for advanced therapeutic applications.

The turn around time for circular RNA production is 3 weeks. Overall turn around time for circular RNA from gene synthesis to circular RNA will be about 7 weeks.

We support insert sequences upto 7Kb. For custom lengths, please contact our technical team.

Yes, we offer T4-based ligation methods to ensure completely scarless circular RNA.

Our QC test for circular RNA include agarose gel electrophoresis assay, PH, endotoxin assay. We have add on QC for testing the circular RNA purity by HPLC method, or RNaseR digestion assay as identification assay.

Yes, modifications are available such as 5% M6A. Please consult our technical team for further customizations.

Additional mRNA Offerings

Custom mRNA

High-quality custom mRNA production via our proprietary IVT RNA manufacturing workflow.

Catalog IVT RNA

Our off-the-shelf mRNA is optimized with 5’ cap and polyA tails that is ready to use for your research needs

Lipid Nanoparticle Packaging

LNP Packaging

mRNA QC Analysis

Send us your in house mRNA samples for expert QC analysis.

Get in Touch withGenScript Custom Circular RNA Synthesis

REQUEST A QUOTE

EMAIL

[email protected]

PHONE

1-877-436-7274

ONLINE FORM

Online Quote Submission

FAX

1-732-210-0262

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Helpful context for this guide

Source-derived material selected through this article’s indexed topics.

Research context

Read sources and limitations before applying a claim.

Case Studies

Fluc mRNA(N1-methyl-pseU) was encapsulated with SM102-LNP or SM102-LNP decorated with anti-Human CD3 Antibody (Clone: OKT3, mAb, Mouse, GenScript, A02199), 100ng of mRNA encapsulated in LNP were incubated with jurkat cells in RPMI140 +10%FBS at 37C for 24 hrs. The expression of Fluc was detected using Luciferase Assay (Promega, E4030). eGFP mRNA(N1-methyl-pseU) was encapsulated with SM102-LNP or SM102-LNP decorated with anti-Human CD3 Antibody (Clone OKT3, mAb, Mouse, GenScript), 10-150ng of mRNA encapsulated in LNP were incubated with primary T cells for 24 hrs. The expression of eGFP was detected using flow cytometry. W1, W2 and W3 indicate different wash conditions. Fluc mRNA(N1-methyl-pseU) was encapsulated with SM102-LNP or SM102-LNP decorated with anti-mouse CD3 Antibody, 0.3mg/kg of mRNA encapsulated in LNP were injection through tail vein into mice, after 24 hrs, mice were sacrificed, and the expression of Fluc mRNA was measured with whole body imaging with different organs. *Not sure which T-cell targeted LNP fits your application? Check our LNP selector tool for guidance >> Resting primary T cells were seeded at 1 × 10⁶ cells per well in a 24-well plate and transfected with 500 ng of generic LNP, CD3Ab-LNP, or CD5Ab-LNP. The anti-human CD3 antibody (OKT3, mAb, mouse, GenScript) and anti-human CD5 antibody (mAb, Rabbit, GenScript) were used for LNP surface decoration. T-cell expression was assessed by flow cytometry (FACS) at 48 h post-transfection. Because CD3Ab (OKT3) is known to activate primary T cells, CD3Ab-LNP exhibited a higher expression level compared with CD5Ab-LNP. Notably, despite the intrinsic difficulty of transfecting resting T cells, CD5Ab-LNP still achieved over 80% transfection-positive cells. Resting primary T cells were seeded at 1 × 10⁶ cells per well in a 24-well plate and transfected with 500 ng of generic LNP, CD3Ab-LNP, CD4Ab-LNP, or CD8Ab-LNP. The anti-human CD3 antibody (OKT3, mAb, mouse, GenScript), anti-human CD4 antibody (F001, mAb, mouse, GenScript), and anti-human CD8 antibody (F002, mAb, mouse, GenScript) were used for LNP surface decoration. T-cell expression was assessed by flow cytometry (FACS) at 48 h post-transfection. CD4Ab-LNP and CD8Ab-LNP exhibited selective delivery to CD4⁺ and CD8⁺ T cells, respectively, with minimal off-target expression. EGFP mRNA(N1-methyl-pseU) was encapsulated with SM102-LNP or SM102-LNP decorated with anti-Human CD34 Antibody (Purified anti-human CD34 antibody(Anti-Human CD34 Antibody, mAb, Mouse, GenScript, CP0001) 100ng of mRNA encapsulated in LNP were incubated with Kasumi-1 cells for 24 hrs. The expression of eGFP was detected using flow cytometry. EGFP mRNA(N1-methyl-pseU) was encapsulated with ALC0315-LNP or ALC0315-LNP decorated with anti-Human CD34 Antibody (Purified anti-human CD34 antibody(Anti-Human CD34 Antibody, mAb, Mouse, GenScript, CP0001) 100ng of mRNA encapsulated in LNP were incubated with primary human CD34+ cells for 24 hrs. The expression of eGFP was detected using flow cytometry. Fluc mRNA(N1-methyl-pseU) was encapsulated with LNP , 0.3mg/kg of mRNA encapsulated in LNP were injection through tail vein into mice, after 24 hrs, mice were sacrificed, and the expression of Fluc mRNA was measured with whole body imaging with different organs. EGFP mRNA(N1-methyl-pseU) was encapsulated with SM102-LNP or SM102-LNP decorated with TriGalNac, 0.3mg/kg of mRNA encapsulated in LNP were injection through tail vein into mice, after 24 hrs, mice were sacrificed, and liver organs were lysed. The eGFP expression in lysate was detected using western blot method. Fluc mRNA (m1Ψ) was encapsulated with LNP, 0.3mg/kg of mRNA encapsulated in LNP were injection through tail vein into mice. Conjugation of anti-CD31 antibodies to LNPs can enhance mRNA expression levels in the lungs of mice. FACS analysis of lung cells revealed that CD31-Ab/LNPs exhibit clear targeting specificity toward CD31⁺ cells. Fluc mRNA (m1Ψ) was encapsulated with LNP, 0.3mg/kg of mRNA encapsulated in LNP were injection through tail vein into mice. Mannose/LNP improved Fluc mRNA expression in spleen and lymph node system. Mannose/LNPs delivering sgRNA targeting Zeb2 achieved efficient gene editing in Cas9⁺ BMDM cells and significantly suppressed tumors in a Cas9-GFP mouse model. Fluc mRNA (m1Ψ) was encapsulated with LNP, 0.3mg/kg of mRNA encapsulated in LNP were injection through tail vein into mice. KKEEE/SM102 LNP enhanced Fluc mRNA expression in kidney organ ~4.6 times comparing undecorated LNP.

Source: genscript.com ↗

Case Studies

1 SM102-LNP espCas9 TRAC HEK293T 97%, Jurkat 82%, T cells72% , HepG2 86% 2 ALC0315-LNP HEK293T 87%, Jurkat 80%, T cells 61% 3 LP01-LNP HEK293T 63%, Jurkat 75% 4 20DODAP-LNP PCSK9 HepG2 87% 5 SM102-SOPC LNP HepG2 90% 6 HifiCas9 HEK293T 97%, T cells 61% , HepG2 88% 7 8 SM102 LNP Cas12a Ultra HEK293T 63% 9 SM102-Mod LNP PEmax HEK3 pegRNA Nicking guide RNA HEK293T 62% 10 CXM2-LNP CBE HEK293T 34% 11 HEK293T 42% ReadyEdit LNP formulations demonstrate robust delivery of prime editing PE6g mRNA and HEK3 pegRNA across multiple cell types. Using optimized lipid compositions, CXM-2 LNP and SM102-Mod LNP achieved editing efficiencies of up to 80% in HEK293T cells, 36% in HepG2 cells, and 43.5% in Jurkat cells. These results highlight that CXM-2 LNP and SM102-Mod LNP enable efficient prime editing in cells. A breakthrough co-delivery platform using two lipid nanoparticles (LNPs): One delivers Cas9 mRNA + sgRNA for precise cutting at RAB11 The other delivers HDR donor template (circular-ssDNA) optimized for stability & integration Advantages: Achieves high-efficiency knock-in Enables robust tagging, reporter insertion, or functional studies at the endogenous RAB11 locus Scalable, non-viral, and suitable for both in vitro and in vivo applications Dual-LNP Delivery System for Highly Efficient Cas9 mRNA-Based Gene Knock-In at the RAB11 Locus Method: SM102-LNP1 loaded with Cas9mRNA and RAB11gRNA, SM102-LNP2 loaded with EGFP coding css-DNA or linear ss-DNA were co-delivered into HEK293T cells at different doses, the knock in efficiency were measured by flow cytometry after 72hrs . Result: higher knock in efficiency was observed using 200ng of circular ss-donor DNA Result: More than 80% editing observed at the TRAC locus Method: espCas9-mRNA : SafeEdit TRAC sgRNA were co-encapsulated in LNP at a molar ratio of 1:10 , 2.4 ug of RNA loaded LNP were incubated with cells for 72 hours, the gene editing efficiency for knocking out TRAC locus was sanger sequenced and editing efficiency was analyzed by ICE Analysis tool Software Result: More than 70% editing and 80% editing observed in 3A9 and Jurkat Cells Method: espCas9-mRNA : SafeEdit TRAC sgRNA were co-encapsulated in LNP at a molar ratio of 1:10 , 2.4 ug of RNA loaded LNP were incubated with cells for 72 hours, the gene editing efficiency for knocking out TRAC locus was sanger sequenced and editing efficiency was analyzed by ICE Analysis tool Software. sgRNA-1 to sgRNA-6 were designed to target different spacer sequence in TRAC gene in mice. Result: 27% knock in observed with the LNP formulation Result: 80% editing efficiency observed in HEK Cells Method: Cas12a mRNA/ SafeEdit TRAC sgRNA (GenScript) were packaged in LNP at 5:1 mass ratio and a serum dependent effect on editing was observed, RNA with SM102-LNP or ALC0315-LNP formulation were incubated with HEK293T cells, cell were lysed at day3 for NGS

Source: genscript.com ↗

About the author

Peptide Therapy Guide Editorial Team

Editorial team for Peptide Therapy Guide.

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Custom Circular RNA Synthesis

For protein coding and non-coding RNA

Custom Circular RNA (circRNA) Service- Scarless circRNA and circRNA with Scar

Benefits of circRNA synthesis with GenScript

Enabling Precision and Efficiency – Scarless circRNA and circRNA with Scar

Custom circRNA Synthesis Workflow: From Design to Delivery

QC Section

Case Study

Popular off-the-shelf circRNA catalogs available now!

Resources

FAQ-Circular RNA Synthesis

Additional mRNA Offerings

Custom mRNA

Catalog IVT RNA

Lipid Nanoparticle Packaging

mRNA QC Analysis

Get in Touch withGenScript Custom Circular RNA Synthesis

REQUEST A QUOTE

EMAIL

PHONE

ONLINE FORM

FAX

Case Studies

Case Studies

Peptide Therapy Guide Editorial Team

IVT RNA Synthesis Services

Targeted LNP Formulation

ReadyEdit LNP Formulation

LNP Formulation