GenCircle Double-Stranded DNA

Applications Field

Cell Therapy

Transposons vector
CRISPR KI template

Gene Therapy

AAV/LV packaging plasmid
non-viral GOI vector
mRNA preparation template

Vaccine

Antigen expression vector
mRNA preparation template

Disease Model
Gene Function Study

Transposons vector
CRISPR KI template

Advantages

Safer! No Antibiotic Resistance Gene

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• No antibiotic resistance gene & antibiotic residual
• Prevent plasmid replication outside of the engineered production host strain

Simpler! Lower Cytotoxicity & Immunogenicity

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• Eliminate CpG islands and other bacterial origin sequences
• Increase stability for longer persistence

Smaller! High fidelity 429bp backbone  

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• Higher gene expression from equivalent plasmid quantity
• Achieve higher transfection efficiency

More Efficient! Increase KI/viral packaging/GOI expression efficiency

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• Increase knock-in efficiency by up to 30%
• Improve viral titer up to 3 folds
• Increase transcription levels up to 4 folds, and protein expression levels up to 39%

Service Details

* For other GOI length & quantity request, please email [email protected] or call 1-877-436-7274.

Service options include：
1. GenScript provides GOI gene synthesis service, and then provides GenCircle dsDNA preparation service.
2. Customer provides GOI in plasmid, then GenScript provides GenCircle dsDNA preparation service.

QC Standards

Case Studies

• Case 1 - GenCircle dsDNA as knock-in template

  Result: KI efficiency increased up to 30% compared to standard plasmid

  • CRISPR KI template（ 2.5 kb insertion）
  
  • Transposon vector for KI（4.8 kb insertion）
  

• Case 2 - GenCircle dsDNA as viral packaging plasmid

  Result: Viral titer improve up to 3 fold compared to standard plasmid, with stable ITR.

  • Lentivirus packaging
  

  Name	Virus Titer (IFU/ml)	Antibiotic resistance gene
  pRRL-PGK-EGFP （Regular Plasmid）	1.23E+8	Significant residue (Ct value<30)
  pRRL-PGK-EGFP MF （GenCircle dsDNA）	3.81E+8	Undetectable (Ct≥35 )

  • AAV packaging
  

  Name	Virus Titer (IFU/ml)	Antibiotic resistance gene
  CMV EGFP AAV2 (Regular Plasmid)	1.12E+12	2%
  CMV EGFP MF (GenCircle dsDNA )	1.22E+12	Undetectable

• Case 3 - GenCircle dsDNA used as GOI nonviral vector

  Result: RNA transcription level increased up to 4 fold, with reduced immune response compared to standard plasmid.

  

• Case 4 - GenCircle dsDNA used as GOI nonviral vector

  Result: protein expression level increased up to 39% compare to standard plasmid.

  

  

Successful Delivered Case

GenScript has successfully delivered multiple orders with a wide range of GOI lengths, including multiple complex or difficult-to-synthesize projects.