Part:BBa_K4586004

Cas12k

Part Description

This part codes for the type V-K CRISPR-Cas system effector protein Cas12k, that is characterized by naturally inactivated RuvC domain and associated with Tn7-like transposon for RNA-guided DNA or genome editing, Its protospacer adjacent motif (PAM) sequence is GGTT. that is essential for activating the enzymatic activity of the Cas protein, and the guide RNA is designed based on the PAM sequence site corresponding to the target gene.

Usage

This part is implemented in our design to be the main element of our therapeutic agent (Cargo) to knock-out the BAFF-R gene apoptosis, which is essential for B-cell survival and maintenance, to cause apoptosis in the autoreactive B-cells that secrete Anti-citrullinated Protein Antibody (ACPA). Its guide RNA is designed to drive the protein portion of the CRISPR Cas system Cas12k to the BAFF-R gene as shown in figure 1.

Figure 1: This figure illustrates the activity of the Cas12k protein into the target autoreactive B-cells through knocking down the B-cell activating factor receptor (BAFF-R) gene that is responsible for B-cells survival and proliferation leading to their apoptosis.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]