The TelN/tos-assisted precise targeting of chromosome segments (TAPE)

Phage N15-Based Vectors for Gene Cloning and Expression in Bacteria and Mammalian Cells

“…Although the techniques to construct linear vectors derived from actual human chromosomes, termed human artificial chromosome (HAC), exist, , the construction method is very laborious and not applicable to existing BAC libraries . A recent example linear cloning platform based on the TelN- tos module, named TelN/tos-assisted precise targeting of chromosome segments (TAPE), has emerged to allow direct cloning of large prokaryotic and eukaryotic genomes without the need to purchase genome sequences from companies or conducting lab synthesis . Briefly, TAPE utilizes the homology arms to clone the target genomic segment into the TelN-linearized plasmid in vitro , followed by the transformation of the resulting plasmid into the TelN-expressing E. coli for vector propagation .…”

“…14 A recent example linear cloning platform based on the TelN-tos module, named TelN/tos-assisted precise targeting of chromosome segments (TAPE), has emerged to allow direct cloning of large prokaryotic and eukaryotic genomes without the need to purchase genome sequences from companies or conducting lab synthesis. 45 Briefly, TAPE utilizes the homology arms to clone the target genomic segment into the TelN-linearized plasmid in vitro, followed by the transformation of the resulting plasmid into the TelNexpressing E. coli for vector propagation. 45 This strategy was shown to have effective cloning of 156 kb bacterial choromosomal fragment, 124 kb yeast genomic fragment, and 16 kb mouse mitochondrial fragment without being limited by the homologous sequences of E. coli host's genome, 45 proving the TelN-tos module is a great prospect for intricate genetic manipulations and cloning procedures.…”

“…45 Briefly, TAPE utilizes the homology arms to clone the target genomic segment into the TelN-linearized plasmid in vitro, followed by the transformation of the resulting plasmid into the TelNexpressing E. coli for vector propagation. 45 This strategy was shown to have effective cloning of 156 kb bacterial choromosomal fragment, 124 kb yeast genomic fragment, and 16 kb mouse mitochondrial fragment without being limited by the homologous sequences of E. coli host's genome, 45 proving the TelN-tos module is a great prospect for intricate genetic manipulations and cloning procedures.…”

“…There is growing evidence to support the rationale that the N15-based linear cloning vector is preferred over standard circular plasmid to circumvent the cloning of huge, unstable repetitive sequences, or abnormal DNA sequences commonly existing in nature, including the production of viral vectors , and cloning of human genes. , The high cloning fidelity of N15-based linear vectors may provide the simpler alternatives for several genetic studies: (i) discovery of novel genes involved in human diseases; (ii) assembly of a more uniform (decreased size bias) DNA libraries containing larger fragments; (iii) complete finishing of large genome sequencing by allowing closure of sequencing gap caused by Sanger or next-generation sequencing; and (iv) establishment of cell culture or animal disease models related to large repetitive sequences. For instance, the N15-based linear vector was used to generate stable cDNA clones of the Zika virus (ZIKV) genome with inherent instability, allowing the production of recombinant ZIKV in transfected Vero cells for the study of ZIKV pathogenicity in mouse model .…”

Breakthrough in efficient cloning and activation of large cryptic biosynthetic gene clusters from high GC actinobacteria

“…For example, CATCH technology based on CRISPR/Cas9 and Gibson assembly [ 4 ], ExoCET technology formed by integrating CRISPR/Cas9 and RecET [ 5 ], CAPTURE technology using CRISPR/Cas12a and loxP site recombination [ 6 ], etc. In addition, TAPE technology based on TelN/tos and Gibson assembly is also a facile and efficient method for large fragment assembly [ 7 ].…”