| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | e chose gene fragments encoding complete deaminase domains as well as extra N and C protein sequences for commercial synthesis (GenScript)/The DdCBE vectors including NLS, TALE array sequences, candidate cytidine deaminases, and UGI sequence were codon optimized for both human and mouse, synthesized commercially (Genscript), and cloned into pCMV_BE4max vector (Addgene#112093),yielding vectors with CMV::NLS-TALE-deaminase-UGI-NLS::bGH expression cassettes./To construct AAV vectors, the sequences between ITRs were synthesized (GenScript) and cloned into pX601 vector (Addgene#61591), followed by sgRNA target sequence cloning steps | Get A Quote |
he elucidation of protein function and its exploitation in bioengineering have greatly advanced the life sciences. Protein mining efforts generally rely on amino acid sequences rather than protein structures. We describe here the use of AlphaFold2 to predict and subsequently cluster an entire protein family based on predicted structure similarities. We selected deaminase proteins to analyze and identified many previously unknown properties. We were surprised to find that most proteins in the DddA-like clade were not doublestranded DNA deaminases. We engineered the smallest single-strand-specific cytidine deaminase, enabling efficient cytosine base editor (CBE) to be packaged into a single adeno-associated vir... More
