A：1. sgRNA Design. 2. sgRNA Synthesis.3. Library construction and plasmid preparation. 4.NGS Sequencing. 5. Virus Packaging (optional). To charge according to this step, we need to first confirm with the client whether they require designing? If yes, please provide the species and gene ID. If no, please provide the sgRNA sequences and library vector sequence then we will evaluate the price.

A：

Single-vector system
The guide RNA (gRNA) and the genome-editing/regulatory effector are encoded on the same plasmid. Delivering this single plasmid enables editing or transcriptional regulation.

Dual-vector system
The gRNA and the effector are carried on two separate plasmids; co-delivery is required for function.

Libraries

• Knockout library: gRNAs designed to generate geneloss-of-function across targets.

• Activation library: gRNAs that, together with aDNA-binding effector fused to activation domains,up-regulate target gene transcription.

• Inhibition (repression) library: gRNAs that, together with aDNA-binding effector fused to repression domains,down-regulate target gene transcription.

We provideboth single- and dual-vector formats for knockout and activation libraries; theinhibition library is available in asingle-vector format.

A：No, gene activation and inhibition generally target different regions of the promoter, knocking out the library targets the CDS region.

A：The concentration should ideally reach 200ng/ul.

At present, most of the products on the market use human genome sgRNA libraries. The screening results obtained from genome sgRNA libraries are more broad-spectrum, but the experimental engineering is large. And Syno human pathway library has the following characteristics：

1、The total number of genes is small, and the storage capacity of the sgRNA library is small, which reduces the amount and production time of experimental engineering.

2、The use of Syno human derived sgRNA libraries has strong specificity, usually through target screening within a known gene range.

3、Multiple pathways can be selected for simultaneous research.

Therefore, before constructing using a Syno human source sgRNA library, it is important to understand the research objectives and pathway genes, and select a single pathway or a combination of multiple pathway sgRNA libraries according to the needs.

Yes, Synbio Technologies offers design services for sgRNA libraries to target both a single gene as well as an entire genome.

Our customers need to provide either the target sequence, the gene name or genome information in order for Synbio Technologies to provide sgRNA design services.

The turnaround time will vary depending on the size of the library and the chosen quality control methods used. The turnaround time for this service starts at nine to thirteen weeks.