{"id":11534,"date":"2025-03-17T12:50:26","date_gmt":"2025-03-17T12:50:26","guid":{"rendered":"https:\/\/www.caltagmedsystems.co.uk\/information\/?p=11534"},"modified":"2025-03-18T15:53:55","modified_gmt":"2025-03-18T15:53:55","slug":"accubase-base-editor","status":"publish","type":"post","link":"https:\/\/www.caltagmedsystems.co.uk\/information\/accubase-base-editor\/","title":{"rendered":"KACTUS: AccuBase\u00ae Base Editor"},"content":{"rendered":"\n

Engineered base editor with near-zero off-target effects<\/h2>\n\n\n\n

AccuBase\u00ae<\/a>\u00a0is a cytosine base editor that converts a C\u2022G base pair into a T\u2022A base pair in the genome. It is independently designed by Base Therapeutics and manufactured for global sales by KACTUS. It creatively embeds the deaminase inside the Cas protein to prevent random tethering of deaminase to non-target site, significantly reducing off-target occurrence while still maintaining high editing efficiency.<\/p>\n\n\n\n

Leveraging their SAMS\u2122<\/a>\u00a0protein engineering and expression platform, KACTUS has developed a large-scale production process for\u00a0AccuBase\u00ae\u00a0base editor. They have focused on maximising the stability, purity, and activity of their GMP-grade\u00a0AccuBase\u00ae, ensuring it meets the highest quality standards in compliance with cGMP raw materials for CGT drug manufacturing.<\/p>\n\n\n\n

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View AccuBase\u00ae Products ><\/strong><\/a><\/div>\n<\/div>\n\n\n\n

How does AccuBase\u00ae work?<\/h2>\n\n\n\n

Precision editing by activating only upon accurate target binding<\/h3>\n\n\n\n

After forming a ribonucleoprotein (RNP) with sgRNA, the\u00a0AccuBase\u00ae\u00a0protein remains in a non-editing state before binding to the target DNA. The deaminase is encapsulated inside the Cas9n protein and does not interact with any non-target DNA, thus significantly reduces the risk of off-target effects. When the RNP binds to the target DNA, the conformation of\u00a0AccuBase\u00ae\u00a0changes, leading to the exposure of the deaminase domain and effectively editing bases within the 3-12 window range of the target site (with the first position being the farthest from the PAM) (Figure 1).<\/p>\n\n\n\n

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Figure 1. AccuBase\u00ae in the non-editing stage (left) and editing stage (right).<\/figcaption><\/figure>\n\n\n\n

Mechanism of action<\/h3>\n\n\n\n

The mechanism of action of\u00a0AccuBase\u00ae\u00a0involves the precise editing of target DNA sequences within the 3-12 base window of the target site, leading to specific point mutations (C to T, G to A) or gene knockouts through the introduction of stop codons or alteration of splicing sites (Figure 2).<\/p>\n\n\n\n

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Figure 2. The gene editing mechanism of\u00a0AccuBase\u00ae.<\/figcaption><\/figure>\n\n\n\n

AccuBase\u00ae\u00a0is ideally suited for correcting a single point mutation or for gene knockouts by introducing premature stop codons or inducing exon skipping by disrupting splice acceptor (SA) sites or splice donor (SD) sites.<\/p>\n\n\n\n

Product Performance Data<\/h2>\n\n\n\n
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Near-zero off-target effects<\/h3>\n\n\n\n

High editing activity<\/h3>\n\n\n\n
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Figure 3. Measurement of off-target effects by GOTI. By leveraging the GOTI (genome-wide off-target analysis by two-cell embryo injection) to measure the off-target effects throughout the whole genome, it was shown that compared to the control base editor (with 700 SNVs detected), the number of SNV obtained after editing by AccuBase\u00ae is similar to the GFP (negative control) group, suggesting a near-to-zero off-target effects of AccuBase\u00ae.<\/p>\n\n\n\n

Figure 4. AccuBase\u2122 RNP was electroporated into activated primary T cells. According to flow cytometry, AccuBase\u2122 can efficiently knock out PD1, B2M, and TRAC proteins on the membrane of activated primary T cells at the protein level. For PD1 and B2M, the knockout efficiency exceeded 80%, while the knockout efficiency of TRAC reached 96%. FACS efficacy % = (positive control – KO group)\/positive control* 100%.<\/p>\n\n\n\n

Avoids INDEL creation<\/h3>\n\n\n\n

Avoids chromosomal translocation<\/h3>\n\n\n\n
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Figure 5. Compared with Cas9 protein, AccuBase\u2122 does not produce DNA horizontal insertions and deletions (INDELs).<\/p>\n\n\n\n

Figure 6. Compared with Cas9 protein, AccuBase\u00ae does not cause chromosomal translocation.<\/p>\n<\/div>\n\n\n\n

Product Features<\/h2>\n\n\n\n

KACTUS offers both\u00a0GMP-grade\u00a0and\u00a0research-grade\u00a0AccuBase\u00ae as off-the-shelf catalog products. Their GMP AccuBase\u00ae Base Editor is free from animal-derived components, has greater than 80% purity, and comes with comprehensive quality control documentation as well as additional batch production records upon request. KACTUS are dedicated to supporting your applications of AccuBase\u00ae from preclinical research through commercial manufacturing, backed by our robust technical and quality support.

Learn more about their GMP quality management systems \u2192<\/a>.<\/p>\n\n\n\n

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Manufactured in a GMP-compliant facility<\/p>\n\n\n\n

Raw materials free from animal-derived components<\/p>\n\n\n\n

High purity, activity, and stability<\/p>\n\n\n\n

Traceable documentation for regulatory support<\/p>\n\n\n\n

Off-the-shelf catalog product<\/p>\n<\/div>\n\n\n\n

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View AccuBase\u00ae Products ><\/strong><\/a><\/div>\n<\/div>\n\n\n\n

Product Specifications & Quality<\/h2>\n\n\n\n

Product Specifications<\/h3>\n\n\n\n

Both their GMP-grade\u00a0and\u00a0research-grade\u00a0AccuBase\u00ae adhere to the following product specifications:<\/p>\n\n\n\n

Parameter<\/th>Specification<\/th><\/tr><\/thead>
Express System<\/td>E. Coli<\/td><\/tr>
Concentration<\/td>10mg\/ml<\/td><\/tr>
Molecular Weight<\/td>210.14kDa<\/td><\/tr>
Form<\/td>Liquid<\/td><\/tr>
Storage Buffer<\/td>30 mM Tris, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol, pH8.0<\/td><\/tr>
Storage<\/td>Store at -80 \u00b110\u00b0C<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n

Quality Control Criteria<\/h3>\n\n\n\n

Their GMP-grade\u00a0AccuBase\u00ae meets the following quality control criteria:<\/p>\n\n\n\n

Assay<\/th>Acceptance Criteria<\/th><\/tr><\/thead>
pH<\/td>8.0\u00b10.5<\/td><\/tr>
Concentration<\/td>9.0-11.0mg\/mL<\/td><\/tr>
Purity (electrophoresis)<\/td>\u2265 80.0%<\/td><\/tr>
Purity (RP-HPLC)<\/td>\u2265 88.0%<\/td><\/tr>
Purity (SEC-HPLC)<\/td>\u2265 80.0%<\/td><\/tr>
Residual DNase<\/td>Sample\/Control \u2264 3.0<\/td><\/tr>
Residual RNase<\/td>Sample\/Control \u2264 3.0<\/td><\/tr>
Residual Host Cell Protein<\/td>\u2264 100.0ng\/mL<\/td><\/tr>
Residual Host Cell DNA<\/td>\u2264 200.0ng\/mL<\/td><\/tr>
Endotoxin<\/td>\u2264 10.0EU\/mg<\/td><\/tr>
Sterility<\/td>Negative<\/td><\/tr>
Mycoplasma<\/td>Negative<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n

GMP Compliance<\/h2>\n\n\n\n

GMP-Grade AccuBase\u00ae\u00a0is produced following cGMP standards, ensuring traceability of raw materials, manufacturing processes, and QC release testing. The GMP-Grade AccuBase\u00ae is available in long-term bulk supply with high batch-to-batch consistency to ensure suitability for industrial applications. We also offer a\u00a0research-grade\u00a0version for a seamless transition from preclinical study to clinical\/commercial CGT drug manufacturing.<\/p>\n\n\n\n

Documentation Package<\/h3>\n\n\n\n

GMP-Grade AccuBase\u00ae\u00a0comes with customisable regulatory documentation including Data Sheet, MSDS, COA, TSE\/BSE statement, CoO. KACTUS has an integrative documentation system consisting of a digital quality management system, specification program, criteria support documents, and quality reports. The documentation package is customisable based on relevant regulatory filings.<\/p>\n\n\n\n

Quality Management System<\/h3>\n\n\n\n