Stellate DNA provides a novel class of high quality, sequence verified, endotoxin-free DNA donor (Genome Catalyst or Gatalyst™) as templates for CRISPR-mediated gene integration (Knock-In or KI). The Stellate Gatalyst™ donor is in a proprietary construction of a circular form of single strand DNA circular single-stranded DNA (cssDNA), which stimulates integration by homologous repair (HR) at the correct integration sites while minimizing the non-homologous integration of the donor into other genome loci or error prone repair via non-homologous end-joining.
The benefits of using Gatalyst™ donors include higher targeted integration efficiency, lower cellular toxicity and reduced frequencies of random integration. Gatalyst™ also address an increasing demand for longer payload sizes in KI gene editing (>2 kilobases), which cannot be addressed by the conventional single strand DNA donors for homology driven repair (HDR). The circular formation of the ssDNA donor is also supposed to protect from degradation, which ensures a longer half-life and availability for targeted integration in the cell when compared with the linear ssDNA form.
We synthesize and subclone or take existing double stranded DNA plasmid containing the desired HDR insertion with the homologous arms from our customer and convert the HDR payloads into the Stellate Gatalyst™ clonal production constructs. We will then deliver the high quality, sequencing verified cssDNA donors as the service product. The Stellate DNA Gatalyst™ platform is flexible to generate your preferred ssDNA donor the same way as you design plasmid DNA. This new generation of gene editing donor production system will advance the gene editing field into an efficient, highly cost-effective, high throughput era that can expedite cell engineering, animal model generation and cancer therapeutic research.
· Largest cargo capabilities (up to ~20 kb)
· High quality without any contamination of dsDNA and RNA
· Full length verified by sequencing
· Minimal Endotoxin levels
· Scalable amount of HDR donor (>100 micrograms)
A260/A280 1.80 – 2.00
Appearance Clear and colorless
Identity Gel Analysis / DNA sequencing verification
dsDNA/RNA contamination Not visible on agarose gel
Endotoxin <1.0 EU/mg
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Iyer S et al., Efficient Homology-directed Repair with Circular ssDNA Donors. bioRxiv doi: https://doi.org/10.1101/864199 (2019).
Li, H et al., Design and specificity of long ssDNA donors for CRISPR-based knock-in. bioRxiv doi: https://doi.org/10.1101/178905 (2019).
Roth, T.L et al., Reprogramming human T cell function and specificity with non-viral genome targeting. Nat. Lett. 559, 405–409 (2018).