CRISPR/Cas12b System Experiment

NegotiableUpdate on 03/04

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Nature of the Manufacturer: Producers

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Online Consult

Overview

CRISPR/Cas12b System Experiment: CRISPR/Cas12b (formerly known as C2c1) belongs to the Class 2 Type V-B CRISPR system and is an important gene editing tool after Cas9 and Cas12a.

Product Details

1、 Molecular mechanism and structural characteristics

（1） System definition and classification

CRISPR/Cas12b(Originally known asC2c1）Belonging toClass 2 Type V-B CRISPR SystemIt is an important gene editing tool after Cas9 and Cas12a. Its core features include:

Double RNA guidance mechanism：

Need tocrRNAWithtracrRNA(Can be fused into sgRNA) jointly guides targeted cleavage, combining some characteristics of Cas9 and Cas12a.

cutting mode：

SingleRuvC structural domainCut double stranded DNA to form5-6 nt viscous end(5 'protrusion), conducive to homologous recombination repair (HDR).

PAM identification：

DependencyT-rich PAM sequence(such as 5 '- TTn-3'), expands the editing ability of AT enriched genomes.

（2） Structural advantages

feature	Cas12b	Compare Cas9/Cas12a
Protein size	Approximately 1100 amino acids(Minimum)	Smaller than Cas9 (1600 aa) and Cas12a (1300 aa)
RNA demand	CrRNA+tracrRNA (can be fused into sgRNA) is required	More complex than Cas12a (requiring only crRNA), but simpler than Cas9
thermal stability	Temperature sensitive type(Activated at 40-55 ℃)	Engineering modification is required to adapt to mammalian cells

2、 Technical advantages and performance

（1） Core advantages

Low off target rate：

GUIDE seq detection shows off target rateSignificantly lower than Cas9(Error insertion/missing reduced by>50%).

high specificity：

CrRNA guidance sequences are longer (>42 nt) and have higher targeting accuracy.

Edit Mode：

InducingLow pure insertion mutation rateMore inclined to generate small fragment deletions, reducing functional unpredictability.

Virus vector adaptability：

Small molecular weight makes it easier to packageAAV or lentiviral vectorImprove the efficiency of in vivo delivery.

（2） Comparison with similar systems

parameter	Cas12b	Cas9	Cas12a
PAM sequence	5 '- TTn-3' (T enrichment)	5 '- NGG-3' (G enrichment)	5 '- TTTV-3' (T enrichment)
Cutting end	黏性末端（5' 突出）	Flat end	黏性末端（5' 突出）
Trans cutting activity	❌ none	❌ none	✅ Yes (capable of cleaving ssDNA)
polygenic editing	More sgRNA is needed	More sgRNA is needed	✅ Support crRNA array

noteCas12b cutting mode enables it togene therapyTo avoid the generation of new epitopes and reduce immune risks.

3、 Application scenarios and cutting-edge cases

（1） Biomedical Applications

antiviral therapy：

HIVSingle editing in T cells can eliminate all infectious HIV, with higher efficiency than Cas9.

Genetic disease repair：

Joint HDR pathway implementationPrecise repair of β - thalassemia geneHemoglobin returns to normal levels.

tumor immunology：

EditPD-1/CTLA-4 dual targetsReduce CAR-T cell exhaustion.