CRISPR crRNA/tracrRNA/gRNA Synthesis Services
CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats) is a newly emerging biotechnology, with sgRNA directing Cas nucleases to perform specific DNA modifications for targeted genes.
Work diagram of CRISPR / Cas9
Creative Biolabs is a professional biotechnology services platform, provide outsourced experiments for oligonucleotide synthesis research. We have professional researchers and strict quality control to ensure the synthesis of high-quality CRISPR crRNA/tracrRNA/Cas9 mRNA services.
Interaction mechanism of crRNA, tracrRNA and gRNA:
CrRNA (CRISPR-derived RNA) binds to tracrRNA (trans-activated RNA) through base pairing to form the tracrRNA/CrRNA complex. It directs the nuclease Cas9 protein to cut double-stranded DNA at the sequence target paired with crRNA. Two kinds of RNA, crRNA and tracrRNA, are artificially designed to be converted into guiding sgRNA (single guiding RNA), thus directing Cas9's targeted DNA cutting.
Schematic of CRISPR / Cas9 gene editing
Applications of CRISPR crRNA/tracrRNA and gRNA:
- Discover genes related to signaling pathways
- Screen drug targets
- Gene therapy
- Gene knockout
- Gene replacement
- Gene activation
- Disease model construction
Features of CRISPR-Cas9 crRNA/crRNA XT/sgRNA
- CRISPR-Cas9 crRNA
CRISPR-Cas9 crRNA must be used with tracrRNA to form a functional gRNA double stranded body. Suitable for most applications. Contains chemical modifications to prevent cellular ribonuclease degradation.
- Crispr-cas9 crRNA XT
Crispr-cas9 crRNA XT must be used in conjunction with tracrRNA to form functional gRNA double-stranded bodies suitable for challenging experimental conditions (e.g., high nuclease environment or Cas9 mRNA). It contains other chemical modifications compared to crRNA that provide cost-effective options to improve stability.
- Crispr-cas9 sgRNA
Crispr-cas9 sgRNA a single RNA molecule consisting of crRNA and tracrRNA sequences is suitable for challenging experimental conditions (e.g, high-nuclease environments or Cas9 mRNA) containing chemical modifications for maximum stability
Advantages of CRISPR crRNA/tracrRNA/gRNA synthesis services:
- Target multiple targets simultaneously
- High cutting efficiency
- The design principle of base complementation is simple
- RNA, DNA and proteins can be co-located simultaneously
- The recognition was unaffected by genomic methylation
In order to meet different experimental requirements, launch CRISPR/Cas9 technology services, according to your need to tailor your own CRISPR/Cas9 vector, including pCas9 / gRNA1 vector and pCas9 / gRNA3 vector, and can use pCas9 / gRNA1 or pCas9 / gRNA3 vector for gene knockout service.
(1) pCas9 / gRNA1 vector
PCas9 / gRNA1 vector can simultaneously express anthropogenic Cas9 protein and gRNA in mammalian cells, and only transfection of -plasmids is required to achieve the cleavage of target genes.
Cas9 schematic diagram of DNA cutting
PCas9 / gRNA1 vector
(2) pCas9 / gRNA3 vector PCas9 / gRNA3 can simultaneously express humanized Cas9Nickase protein and gRNA in mammalian cells.
Cas9Nicknase schematic diagram of DNA cutting
PCas9 / gRNA3 vector
Features of Cas9Nicknase:
- D10A mutant of Cas9 protein
- DNA cutting single strand
- Nick gap is quickly repaired by cells, and few gene mutations occur
- gRNA pairs are needed to realize DNA double-strand break
- Nicknase protein is specific for gene knockout
References
- Nicholas Sofos, Mingxia Feng, et al. Structures of the Cmr-β Complex Reveal the Regulation of the Immunity Mechanism of Type III-B CRISPR-Cas. Molecular Cell, 2020, 79(5): 741-757.
- Jang Hyeon-Ki, Song Beomjong, et al. Current trends in gene recovery mediated by the CRISPR-Cas system. Experimental & Molecular Medicine, 2020, 52(7).
- Hong-Tu Z, Yan-Li W. Molecular mechanism of crRNA biogenesis and interference in CRISPR-Cas system. Chinese Bulletin of Life ences, 2016.
*For Research Use Only. Not for use in diagnostic procedures.