Difference between revisions of "Part:BBa K2457005"

Revision as of 00:57, 2 November 2017

Standardized pBAD_Cas9 device

This BioBrick is composed by BBa_K2457000 (AraC + pBAD) and BBa_K2457001 (Cas9), where AraC + pBAD regules the Cas9 expression through your repressor and your inductor: Glucose and L-Arabinose, respectively, and Cas9 is a protein that cleaves a target sequence in the genome. This module was developed by the Amazonas_Brazil team to compose the CRISPeasy toolbox, subdivided in CRISPeasy-out and CRISPeasy-in.

CRISPeasy-out is a standardized knock out approach that aims to turn the current method in an easier way to engineer the bacterial genome by Non-Homologous End Joining (NHEJ).

CRISPeasy-in is a standardized knock-in approach that aims to turn the current method in an easier way to insert a donor template into the bacterial genome by Homology-Directed Repair (HDR).

Usage and Biology

Design

Figure 1: BBa K2457005 circuit

Characterization

Sequencing

Figure 2: Sequencing electropherogram from BBa_K2457005.

Figure 3: Alignment of the designed sequence and our final construction from BBa_K2457005.

Sequence and Features