Description
Recombinant Cas13a Protein for Precision RNA Detection and Cleavage, mRNA Knockdown, and Phage Genome Editing Technologies: LbuCas13a – glycerol free formulation
- Recombinant LbuCas13a for forming RNP complexes with guide-RNA
- Highly stable and free of RNAses
- Highly purified and extremely active (RNA cleavage)
- Trans ssRNA cleavage activity can be used to cleave ssRNA reporters generating detectable signals
- Suitable for RNP-transfection of primary cell cultures and high-throughput use of instruments with sensitive fluidics due to glycerol absence
- Bulk Pricing available – please contact us for a quote
SignalChem’s highly purified recombinant Leptotrichia buccalis Cas13a (LbuCas13a) protein (>95% purity) without nuclear localization signal (NLS) is used for CRISPR-based RNA detection technologies and for CRISPR-mediated in vitro applications. The enzyme was expressed in E. coli and is provided tag-free.
LbuCas13a protein combines with guide RNA (gRNA) to form a stable ribonucleoprotein (RNP) complex that enables the inactive nuclease to be guided to the specific RNA target site. Once guided to the specific cleavage site, LbuCas13a becomes activated to specifically and precisely cleave the RNA hrough conserved basic residues within its two HEPN domains, and to indiscriminately trans-cleave unspecific single stranded RNA (ssRNA) nearby. A short single-stranded RNA labeled with a fluorophore and a quencher on the two termini can be used as a reporter to generate a detectable fluorescence signal for highly precise detection of the guide RNA-targeted RNA sequence. The highly precise detection system can be combined with pre-amplification procedures including isothermal amplifications such as LAMP, to achieve one-pot reaction systems, suitable for field-testing, and offering many advantages over qPCR-based detection.
Moreover, Cas13a proteins have been shown to knock down oncogene mRNAs very efficiently and are extremely promising potential precision oncology tools.
LbuCas13a in particular has also found to be very efficient in broad spectrum phage genome editing.
SPECIFICATIONS
PURITY | >95% (SDS-PAGE gel analysis)
ENZYME SOURCE | Leptotrichia buccalis Cas13a is expressed in E. coli without nuclear localization signal (NLS), and is provided tag-free.
QUANTITY | 100 ug, plus 1ml 10x Cas13 Reaction Buffer II
STORAGE/SHIPPING CONCENTRATION | 0.1 ug/ul
STORAGE BUFFER | 50mM Tris, pH7.5, 600mM NaCl, 0.1mM TCEP.
RECOMMENDED STORAGE CONDITIONS | -80°C
EXPIRATION | 12 months from receipt when stored as directed. Avoid repeated freeze/thaw cycles.