CRISPR/Cas9: gRNA Design and Vector Construction (This service is temporarily unavailable)

CRISPR/Cas9 Service: gRNA Design

• Three gRNAs are designed in common exons of different transcriptional products, generally.
• Target positions are as far as possible in the first third of the gene, and targets can destroy important domains of the protein and all the alternatively spliced transcripts.
• Six gRNAs are designed if the effects are not good and the time is pressing.

CRISPR/Cas9 Service: Vector Construction

  • Common Cas9 vectors or lentiviral transfer vectors are constructed according to the customer's needs.
  • Lentiviruses for gene knockout are prepared and delivered to customers.
  • Donor vectors for HDR also are designed and prepared for customers.
  • The knockout efficiency of our vector is very high. For example, using the same gRNA, our knockout efficiency is 64%, while the other company's is 26%.

CRISPR/Cas9 Service Specification

Service contents Provided by customers Delivery time Comments
• Construction of Common Cas9 vectors or lentiviral transfer vectors
The gRNAs slao can be constructed into the customer's vector (or that customer specify).
• Preparition of lentiviruses for gene knockout
• Design and preparion of donor vectors for HDR
Gene names of a species or gene IDs ~2 weeks
(If gRNAs need be validated by cell transfection, it will take a total time of 6-8 weeks depending on the difficulty levels of the transfection.)
Please inquire support staff if gRNAs need be validated by cell transfection.

In order to provide quotation efficiently and accurately, please download and complete the inquiry sheet, then fill out the whole inquiry sheet and email it to us, we will reply to you in time.

Download and complete the request form.
Tel: +86-400-890-9989

CRISPR/Cas9 Service Cases

KO vectors with gRNAs were transfected into cells. After 3 days single colnes with fluorescence were obtained by limited dilution. The genomes of each single clone were extracted to perform PCR and sequencing analysis.

1. Genomic sequencing results of Gln synthysis-related gene KO cell lines

2. Genomic sequencing results of glycosylation-related gene KO cell lines

3. Genomic sequencing results of TCR gene KO cell lines

CRISPR/Cas9 System

Clustered regularly interspaced short palindromic repeats (CRISPR) are segments of prokaryotic DNA containing short repetitions of base sequences. Each repetition is followed by short segments of "spacer DNA" from previous exposures to a bacteriophage virus or plasmid.
The CRISPR/Cas system is an immune system that confers resistance to foreign genetic elements such as viruses and plasmids, and provides a form of acquired immunity in many bacteria and archaea species. The system recognizes and cuts these exogenous genetic elements to prevent repeated attacks in a manner analogous to RNAi in eukaryotic organisms. Bacteria employing CRISPR/Cas systems collect bits of genetic material from viruses or other invaders, and save those sequences as "spacer DNA" in specific regions of their genomes. Those foreign DNA-containing regions are called "clustered regularly interspaced short palindromic repeats(CRISPR)", When the cell undergoes a later attack, the spacer sequences stored among the CRISPR elements are expressed as CRISPR-RNAs (crRNAs). crRNAs binding with trans-activating crRNAs (tracrRNAs) are processed into complexes with CRISPR-associated proteins (Cas proteins). These complexes have the specificity of each spacer sequence. Target double-strand break(DSB) by these complexes requires base pairing between the crRNA and the target DNA. Target recognition is facilitated by the presence of a short sequence called a protospacer adjacent motif (PAM) that conforms to the sequence NGG (spCas9 recongnizes).
CRISPRs are found in approximately 40% of sequenced bacterial genomes and 90% of sequenced archaea.
crRNA contains the RNA used by Cas9 to guide it to the correct section of host DNA with tracrRNA forming an active complex with Cas9.
tracrRNA binds to crRNA and forms an active complex with Cas9.
gRNAs are a combined RNA consisting of a tracrRNA and a crRNA.