Human Artificial Chromosome Building Service

The idea of human artificial chromosome (HAC) has provided new ideas for vector research. Lifeasible offers HAC vector construction services using different strategies to help you use them as gene therapy vectors as soon as possible. Our HAC vectors do not require a viral protein capsid and can exist in a linear or circular form like a natural chromosome. At the same time, in the absence of selective pressure, they can be stabilized in the cell with a low copy number. Has unlimited cloning ability. It can insert target genes and other regulatory elements or specific cell-targeting elements.

Top-down HAC building strategy

We construct new HACs based on broken fragments of natural chromosomes. We can obtain broken fragments by various methods, including low-dose radiation, telomere-localized chromosome truncation technique, amplification of exogenous DNA fragments after targeted insertion, and spontaneous fragmentation of natural chromosomes during mitosis.

• Integrating exogenous telomeric DNA into a mammalian chromosome truncates the chromosome while generating new telomeres at the broken ends. By applying this technique, we could trim mammalian chromosomes and successfully construct. We could insert telomeric DNA into specific sites in the chromosome, and the resulting HACs were structurally stable.
• We can insert exogenous DNA into the mitotic regions of mouse and human chromosomes, and amplifying this DNA results in cleavage of the chromosome into small fragments. These fragments are then purified for microinjection or liposome-mediated transfection to construct HACs.
• We can construct HAC vectors by microcell-mediated chromosome transfer using fragments produced by mitosis of human chromosomes under natural conditions.

Bottom-up HAC building strategy

We construct HACs by assembling mitotic DNA, telomeric DNA, and replication origins either intracellularly. They exist in cells at low copy numbers and exhibit high mitotic stability.

HAC vector expression line construction

We can construct different gene expression systems using HAC. We utilize co-transfection, Cre/loxP, and FLP-FRT systems for site-specific recombination and chromosomal cloning to insert target genes.

Advantages of HAC as a gene therapy vector

• Dividing and replicating with host cell division for stable inheritance.
• Large enough capacity to carry the gene and all its remote regulatory sequences.
• Realization of expression vector and transfer vector into one, eliminating the process of gene transfer to cells and then integration from transfer vector to chromosome before expression. Integrating the target gene and HAC is artificially recombined, and the recombination efficiency can be significantly improved through manual control and screening.
• It does not integrate into the host chromosome and exists independently without causing insertion mutation.
• Avoiding the harm of viral genes in viral vectors to the human body.

If HACs are used as vectors for gene therapy, it will overcome the problems with current gene therapy vector technology. Lifeasible accelerates this process by providing you with HAC building services. If DNA sequences that do not cause aberrant expression are utilized to provide replication initiation sites, there is no doubt that more perfect HACs will be produced, and we are working on this as well. Please feel free to contact us for a solution.