SsDNA COA Synthesis Gene Synthesis Single Stranded DNA Synthesis

SsDNA Synthesis Services Single Stranded DNA Synthesis Gene Synthesis

Recent research in CRISPR/Cas9 technology has shown that the use of long single-stranded DNA (ssDNA) donor templates greatly enhances the efficiency of homology directed repair (HDR) enabling researchers to optimize the process of efficiently generating transgenic animal models and cell lines.

CELLFREE's ssDNA synthesis service provides up to 5000 nt of full sequence-verified fragments quickly and affordably. Our fragments are derived from clonally purified double-stranded DNA (dsDNA), producing the highest quality results possible. We offer services of sequence-verified ssDNA synthesis for CRISPR-mediated gene knock-in, in-vitro transcription, and much more.

ADVANTAGES OF ssDNA SYNTHESIS FOR CRISPR GENE KNOCK-INS

1. Lower cellular toxicity compared to dsDNA after cellular delivery

2. Low off-target integration for more reliable gene knock-ins

3. High specificity knock-in templates for homology directed repair

4. High efficiency donors for targeted insertions and gene replacements

WORKFLOW

Double-stranded breaks are generated through CRISPR/Cas9 editing, then repaired by the endogenous cellular pathways of non-homologous end joining (NHEJ) and HDR. While the HDR pathway has consistently proven successful in copying genetic information via homologous recombination, insertion of exogenous genetic material is a challenge due to the inherent inefficiencies of HDR. Double-stranded DNA has historically been the template of choice for gene insertions, but recent research has shown the superiority of ssODNs as a donor template for HDR. Offering much higher efficiency to insert long sequences with shorter homology arms, ssDNA has become the preferred donor template for this process.

APPLICATIONS OF ssDNA SYNTHESIS

1. Antibody Discovery: Engineer customized cell lines or transgenic mouse models to study in vivo immune responses

2. Food technology: Modify genomes of agricultural crops to study pathogenic resistance for improved food security

3. Cancer Biology: Generate CRISPR gene insertions to study oncogene function for targeted therapeutics

4. Biofuels:  Precise genome editing to optimize metabolic pathways for biofuel production

TURNAROUND TIME 

DELIVERABLES

1. Sequence verified
2. 3 µg, 10 µg, or 20 µg lyophilized DNA
3. Certificate of Analysis (COA) including gel image, sequence trace data with alignment, and sequence files