GenCRISPR™ LbuCas13a Nuclease

GenCRISPR™ LbuCas13a Nuclease is an RNA-guided, RNA-targeting CRISPR enzyme, commonly referred to as LbuC2c2. This Cas13a protein can be applied to detect single RNA molecules with proven high specificity. Recently, Cas13a has been leveraged as a diagnostic nucleic acid detection tool.

价格	￥1600 在线订购包邮(港澳台除外)
产品编号	Z03742-100
规格	100 μg 500 μg 1 mg 大单询价
数量
参考货期


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产品概述应用指导产品属性质控参数图例技术背景

产品概述

Description

GenCRISPR™ LbuCas13a Nuclease is an RNA-guided RNA endonuclease from Leptotrichia buccalis. Cas13a (previously known as C2c2) belongs to the Class 2 Type VI CRISPR-Cas system and contains a single protein effector with two HEPN domains. Cas13a exhibits two distinct ribonuclease activities. Cas13a possesses sequence-specific RNA cleavage activity in vitro and in vivo. And after specific cleavage, Cas13a activates collateral cleavage activity towards adjacent non-specific ssRNA sequences. The two distinct ribonuclease activities of Cas13a enable guide-RNA processing and RNA detection. The RNA detection empowered with Cas13a-crRNA complex and quenched fluorescent ssRNA as reporter has been reported to be versatile, rapid and high-specific.

应用指导

crRNA-dependent RNA cleavage

产品属性

Source	Recombinant Cas13a expressed by E.coli
Species	Leptotrichia buccalis
Tag	Tag-free
Apparent Molecular Weight	~140 kDa, on SDS-PAGE under non-reducing conditions
Concentration	4 mg/ml
Storage Buffer	50 mM Tris-HCI, 600 mM NaCl, 0.1 mM TCEP, 50% glycerol, pH 7.5.
Storage & Stability	Store at -20 °C for 12 months from the date of manufacture. Avoid repeated freeze-thaw cycles. Do not store below -20 °C!
Accession#	C7NBY4

质控参数

Appearance	Clear, colorless liquid
Purity	≥ 90% by SDS-PAGE
Activity	≥ 85% by in vitro digestion of ssRNA
Concentration	4 mg/ml ± 10% as analyzed by A280
Residual DNase	Undetectable
Residual RNase	Undetectable
Endotoxin Level	< 0.2 EU/μg

图例

LbuCas13a has sequence-specific RNA cleavage activity, and after specific cleavage, it activates collateral cleavage activity towards adjacent non-specific ssRNA sequences.

A 50 μL reaction containing ssRNA, Cas13a/crRNA complex, Fluorescent RNA reporter, and Cas13a Reaction Buffer is incubated for 30 minutes at 37 °C. After incubation, the fluorescence intensity is measured using a Microplate Reader (Excitation wavelength: 494 nm; Emission wavelength: 518 nm). »

技术背景

References

1.       Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector." Science 353.6299 (2016).
2.       Gootenberg, Jonathan S., et al. "Nucleic acid detection with CRISPR-Cas13a/C2c2." Science 356.6336 (2017): 438-442.
3.       Gootenberg, Jonathan S., et al. "Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6." Science 360.6387 (2018): 439-444.
4.       Iwasaki, Roman S., and Robert T. Batey. "SPRINT: a Cas13a-based platform for detection of small molecules." Nucleic acids research 48.17 (2020): e101-e101.
5.       East-Seletsky, Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection." Nature 538.7624 (2016): 270-273.

--> 1.       Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector." Science 353.6299 (2016).
2.       Gootenberg, Jonathan S., et al. "Nucleic acid detection with CRISPR-Cas13a/C2c2." Science 356.6336 (2017): 438-442.
3.       Gootenberg, Jonathan S., et al. "Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6." Science 360.6387 (2018): 439-444.
4.       Iwasaki, Roman S., and Robert T. Batey. "SPRINT: a Cas13a-based platform for detection of small molecules." Nucleic acids research 48.17 (2020): e101-e101.
5.       East-Seletsky, Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection." Nature 538.7624 (2016): 270-273.

For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.