Overview

Double-stranded DNA (dsDNA) has been used for CRISPR homology directed repair (HDR) template for creating gene knock-in with high editing efficiency. GenWand™ dsDNA is developed with covalently closed ends to mitigate no-homologous end joining risk and increase knock-in accuracy. GenScript now offers up to g level closed-end dsDNA ideal for screening, process optimization and scale up.

Why closed-end dsDNA as CRISPR Gene Knock-In HDR Templates?

  • Lower toxicity and higher KI efficiency compare to PCR product
  • Closed-end to mitigate NHEJ events
  • No introduction of unnecessary plasmid sequences
  • Ideal for long sequence knock-ins
  • Ideal for large scale screening and scale up
closed-end dsDNA advantage

Why GenWand™ dsDNA Service?

  • Plasmid-based production process
  • 1-10 kb
  • µg to g level production
  • Comprehensive QC to ensure minimal impurities
  • Built in endotoxin removal process and guaranteed < 10 EU/mg Endotoxin
  • cGMP and cGMP-like ssDNA production now available – learn more here

Services

dsDNA

Starting at 50µg/item , Deliver in as fast as 3 weeks .

Length (bp) Price Starting From
1000 $1,200
3000 $2,400
5000 $2,500
10000 $3,000
Test Specifications Detection Method Release Criteria Research Grade
Purity Agarose gel electrophoresis Single band
Sequence accuracy Sanger sequencing 100% sequence alignment
Optical density Spectrophotometer at 260 nm/230 nm ≥ 2.0

GenScript also offers full cGMP and GMP-like ssDNA and dsDNA services enabling faster CAR-T drug development!

Cell Therapy

State-of-the-art facility

Clean suite with class A isolator in a class C background

Gene Therapy

Comprehensive QA/QC & documentation

supporting the IND filing process

Vaccine

All-encompassing non-viral solutions

RUO to cGMP linear ssDNA, dsDNA and miniaturized circular dsDNA

Applications

Mechanism of CRISPR HDR based gene editing

CRISPR based gene insertion, replacement, or correction

Mechanism of CRISPR HDR based gene editing

CRISPR/Cas9 technology is commonly used to create precise double stranded breaks (DSBs) at target DNA sites. The guide RNA (gRNA) recognizes the protospacer adjacent motif (PAM) sequence on the target DNA after forming complex with Cas9, then Cas9 exerts its endonuclease function to cause DSBs. This triggers two mechanisms for repair: one is non-homologous end-joining (NHEJ), which introduces mutations in the DSB site. The other mechanism is homology directed repair (HDR) which enables the donor DNA to be inserted at the break site and create gene knock-ins.

Resources

FAQs

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with GenScript Custom
CRISPR Experts

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