Ready-to-use CRISPR Cas9 sgRNA Synthesis

Instead of conventional time-consuming sgRNA plasmid construction, we offer our worry-free and ready-to-use CRISPR-Cas9 sgRNA synthesis service package to save your time and facilitate your research.

Synbio Technologies sgRNA synthesis service package includes sgRNA target design, DNA template synthesis of sgRNA, sgRNA in vitro transcription, and sgRNA purification to provide customers with high quality ready-to-use sgRNA that can be directly transfected into cells or animals. Synbio Technologies ready-to-use sgRNA saves time on plasmid construction and avoids the drawbacks of potentially non-degraded plasmids.

sgRNAs directly from in vitro transcription have been proven to successfully and effectively perform gene editing in multiple species including zebrafish [1], mouse [2], filamentous fungi [3], etc. Synbio Technologies has also designed 3 universal negative control sgRNA for the Human and Rat genome: Syno®-negative controls sgRNA1, sgRNA2, and sgRNA3. They are fully ready-to-use as negative controls in Human and Rat gene/genome editing experiments.

Competitive Advantages

  • One-stop Solution: Synbio Technologies provides integrated services from target sgRNA design to high purity ready-to-use sgRNA production.
  • Ready-to-use: Directly provide high quality and time-saving sgRNA transcribed in vitro without construct generation.
  • High Quality and Convenient: Ready-to-use sgRNA can be directly transfected into cells, or injected into animals for gene editing study.
  • Fast Delivery: In just 3 business days, Synbio Technologies will deliver up to 20 μg of customized ready-to-use sgRNA.

One-stop Ready-to-use sgRNA Production Workflow

Synbio Technologies has designed a number of sgRNAs to target several genes in mouse, and performing in vitro transfection. The experimental period was shortened to 2 days, and the sgRNA amount was increased to 10-20 μg. This change could means a significant jump in efficiency for synthetic biology experiments utilizing CRISPR-Cas9.

sgRNA Design and DNA Synthesis by PCR

One-step synthesis of DNA template
One-step synthesis of DNA template

sgRNA Construction and Sequence Confirmation

Clone DNA template into pUC57 vector, the sequencing result (Fig. 1) coincided with the designed sequence.

Fig. 1. Comparison between blunt end ligation result and designed sequence

In vitro Transcription

Agarose gel electrophoresis of sgRNA obtained by in vitro transcription, clear bands shown in Fig. 2.


Fig. 2. agarose gel electrophoresis
of sgRNA

sgRNA Verification

Transcript sgRNA into cDNA, design sgRNA amplification primer, and obtain the complementary DNA sequence by PCR reaction. Clone DNA sequence into pUC57 vector; sequencing result (Fig. 3) showed the sgRNA sequence is correct.

*The template of Lane 1 is reverse transcripted cDNA, The template of Lane 2 is sgRNA digested by DNase I; The template of Lane 3 is in vitro transcripted DNA.


Fig. 3. sgRNA sequence verification
using agarose gel electrophoresis

Service Specifications

Service NameProduct/Service SpecificationsTurnaround Time (business day)DeliverablesPrice
Ready-to-use sgRNA synthesissgRNA design DNA template synthesis in vitro sgRNA transcription and purification<10 sgRNA, 5
10-20 sgRNA, 10
>20 sgRNA, Inquiry
sgRNA
COA
Inquiry
Syno® negative control sgRNANegative control sgRNA in vitro sgRNA transcription and purification5Syno®-negative control sgRNA1 Syno®-negative control sgRNA2 Syno®-negative control sgRNA3Inquiry

References

[1] Xiao A, Wang Z, Hu Y, et al. Chromosomal deletions and inversions mediated by TALENs and CRISPR/Cas in zebrafish [J]. Nucleic Acids Res. 2013,41(14):e141.
[2] Fujii W, Kawasaki K, Sugiura K, Naito K. Efficient generation of large-scale genome-modified mice using gRNA and CAS9 endonuclease [J]. Nucleic Acids Res. 2013,41(20):e187.
[3] Liu R, Chen L, Jiang Y, Zhou Z, Zou G. Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system. Cell Discovery. 2015.7.