Large Segment-CRISPR (Leading technology)
Gene editing technology become practical since 2009. It includes three systems ZFN, TALEN and CRISPR/Cas. The most important functional properties of gene editing system are (1) identifiable and cleavable specific DNA sequences and (2) can be performed across species.
Among them, CRISPR/Cas is the easiest and the most efficient system. Through gene editing technology, developers can arbitrarily cut target DNA sequences in any cell and perform subsequent applications. One of the important applications is DNA replacement: when a gene editing proteins such as TALEN or CRISPR/Cas cleaves genomic DNA, a template DNA integrates into the cut site of genomic DNA with the mechanism called homologous recombination. Through DNA replacement, researchers can manipulate cells or organisms to be biofactory that can efficiently produce specific proteins, or to be disease animal models that are important tools for drug development.
The difficulty of DNA replacement increases sharply with the increase of the length of substitution. The technical hurdle is the design and preparation of template DNA.
The average efficiency technical teams perform DNA replacement in rodent with gene editing CRISRP/Cas is as follows:
1~1000 bp, 5~30%;
1000~3000 bp, 1~10%
The limit of DNA replacement for most technical teams is 3000bp.
GEcoll’s frontier technology of large-segment DNA manipulation with CRISPR/Cas allows us to perform DNA replacement of 3000~10000 bp, and the efficiency is between 1% to 5%. We are capable of conducting larger DNA replacement with gene editing.