Recombinant Human AAGAB protein

GGACT recombinant protein is a synthetic fusion protein designed for targeted genome editing applications, particularly in the context of CRISPR-based technologies. The name "GGACT" is derived from its functional domains: a **G**uide RNA-binding protein (dCas9 or Cas9 nickase), a **G**ST affinity tag, **A**denine base editor (ABE), **C**ytidine deaminase (for cytosine base editing), and a **T**ranscriptional activation/repression module. This multifunctional protein integrates key components required for precise DNA modification, enabling simultaneous base editing and transcriptional regulation within a single system.

Developed to address limitations in traditional CRISPR-Cas9 systems, GGACT aims to enhance editing efficiency while minimizing off-target effects. The GST tag facilitates protein purification via affinity chromatography, ensuring high yield and purity for experimental or therapeutic use. The adenine and cytidine deaminase domains allow for C-to-T or A-to-G conversions without inducing double-strand DNA breaks, reducing unintended mutations. Additionally, the transcriptional module enables epigenetic modulation, making GGACT versatile for both gene editing and expression control.

GGACT has been optimized for delivery via viral vectors (e.g., AAV) or lipid nanoparticles, broadening its applicability in vitro and in vivo. Its design reflects advancements in synthetic biology, where modular domains are combined to achieve multifunctionality. Researchers employ GGACT in disease modeling, functional genomics, and potential gene therapies for genetic disorders like sickle cell anemia or cystic fibrosis. Ongoing studies focus on refining its specificity and compatibility with diverse cell types, positioning GGACT as a promising tool for next-generation precision genome engineering.

**Background of AAGAB Recombinant Protein**

AAGAB (alpha- and gamma-adaptin binding protein), also known as p34. is a cytosolic protein implicated in the regulation of clathrin-mediated endocytosis (CME) and intracellular trafficking. It interacts with adaptor protein (AP) complexes, specifically AP-1 and AP-2. which are critical for vesicle formation and cargo sorting. AAGAB contains a conserved N-terminal domain that binds to the γ- or α-adaptin subunits of AP-1 and AP-2. respectively, and a C-terminal region involved in dimerization. This interaction suggests its role in stabilizing AP complexes or modulating their assembly during vesicle budding.

Recombinant AAGAB protein is produced using heterologous expression systems (e.g., *E. coli* or mammalian cells*) to study its structure, biochemical interactions, and functional mechanisms. Its recombinant form enables detailed analyses, such as binding assays with adaptins, structural studies (e.g., X-ray crystallography), and functional experiments in cell-free or cellular systems.

AAGAB gained clinical attention due to its association with punctate palmoplantar keratoderma (PPKP), a rare skin disorder. Mutations in the *AAGAB* gene (e.g., truncations or missense variants) disrupt protein function, leading to abnormal keratinocyte differentiation and epidermal defects. Recombinant AAGAB variants mimicking these mutations have been instrumental in elucidating disease mechanisms, including impaired AP complex interactions and dysregulated epidermal trafficking.

Beyond disease research, recombinant AAGAB serves as a tool to explore CME regulation, membrane trafficking pathways, and adaptor protein dynamics. Its applications extend to drug screening for trafficking-related disorders and engineering synthetic vesicle systems. Overall, AAGAB recombinant protein bridges fundamental cell biology with translational insights into genetic skin diseases.