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Cat. No.: LbuCas13a-100 (for 100pmol)

Cat. No.: LbuCas13a-1k (for 1000pmol)

Description

LbuCas13a nuclease (also known as C2c2) is a crRNA-guided RNA endonuclease derived from Leptotrichia buccalis. In the presence of a protospacer flanking site (PFS) adjacent to the target single-stranded RNA (ssRNA), Cas13a specifically recognizes and cleaves the target RNA.

In addition, Cas13a exhibits collateral (trans) cleavage activity. Once the LbuCas13a protein forms a ternary complex with crRNA and target RNA, it becomes activated and indiscriminately degrades non-specific ssRNA molecules present in the reaction system. This property has been harnessed for the development of rapid nucleic acid detection assays. For example, the Zhang laboratory at the Broad Institute developed the next-generation molecular diagnostic platform “SHERLOCK” based on Cas13a (see PMID: 28408723).

Components

• LbuCas13a Nuclease (10 µM)
• 10× Buffer for Cas13a  

Storage Conditions

• Store all components at –20 °C.
• Transport at ≤0 °C.

Activity Definition

One transU is defined as the amount of Cas13a enzyme required to cleave 1 pmol of ssRNA reporter within 1 minute at 37 °C in a 20 µL reaction system containing 1× Buffer for Cas13a [pH 7.6].

Notes

• Cis-cleavage activity: Guided by crRNA, Cas13a specifically cleaves the target RNA. The requirement for a protospacer flanking site (PFS) on the target RNA is relatively relaxed.
• Trans-cleavage activity: In the presence of target RNA, Cas13a forms a ternary complex with crRNA and the target RNA (Cas13a/crRNA/target RNA). Once activated, Cas13a exhibits trans-cleavage activity, indiscriminately degrading non-specific single-stranded RNA molecules within the reaction system.
• To prevent RNase contamination, maintain a clean and tidy work area. Always wear clean gloves and a mask during operation, and ensure that all consumables (pipette tips, centrifuge tubes, etc.) are RNase-free.
• Cas13a protein is heat-sensitive and prone to inactivation. Reactions should be assembled on ice, and the enzyme should be returned to –20 °C immediately after use.
• The Cas13a protein and reaction buffer provided by the company contain no nuclease activity other than Cas13a.
• For research use only.