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.com
Volume 12
Journal of Molecular and Genetic Medicine
ISSN: 1747-0862
May 21-23, 2018 Barcelona, Spain
&
Integrative Biology
6
th
International Conference on
Genomics and Molecular Biology
10
th
International Conference on
Genomics 2018 and Integrative Biology 2018
May 21-23, 2018
JOINT EVENT
Human artificial chromosomes and TAR cloning technology for genomes studies and biomedicine
Natalya Kouprina
National Cancer Institute-NIH, USA
T
ransformation-associated recombination (TAR) cloning allows selective isolation of full-length genes and genomic loci
as large circular Yeast Artificial Chromosomes (YACs) in yeast. The method has a broad application for structural and
functional genomics, long-range haplotyping, characterization of chromosomal rearrangements and evolutionary studies.
Also, the benefit of combining the TAR gene cloning technology with the HAC gene delivery system for gene expression studies
will be discussed. Human artificial chromosome HAC-based vectors offer a promising system for delivery and expression of
full-length human genes. HACs avoid the limited cloning capacity, lack of copy number control and insertional mutagenesis
due to integration into host chromosomes that plague viral vectors. Recently we engineered the HAC with a single
LoxP
gene
adopter site and a defined structure and demonstrated its utility for delivery of several full-length genes and correction
of genetic deficiencies in human cells. We also showed that phenotypes arising from stable gene expression can be reversed
when cells are “cured” of this HAC by inactivating its kinetochore in proliferating cell populations, a feature that provides a
control for phenotypic changes attributed to expression of HAC-encoded genes, thereby aiding in proper interpretation of gene
function studies. Also, we demonstrated that HAC-bearing ES cells were indistinguishable from their wild-type counterparts:
they retained self-renewal potential and full capacity for multi-lineage differentiation during mouse development, whereas the
HAC itself was mitotically and transcriptionally stable during this process. The HAC vectors have a great potential for genes
function studies, gene therapy, regenerative medicine, screening of anticancer drugs and biotechnology.
kouprinn@mail.nih.govJ Mol Genet Med 2018, Volume 12
DOI: 10.4172/1747-0862-C2-028