Zbornik - Društvo genetičara Srbije

6 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE
Subotica, 30. novembar - 4. decembar 2004.
Uvodno
predavanje
VEŠTAÈKI HROMOZOM BAZIRAN NA SATELITSKOJ DNK
Gyula Hadlaczky
Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary
In vivo konstrukcija animalnog veštaèkog hromozoma pretstavlja ponovljivu i efikasnu
tehnologiju za konstrukciju stabilnog veštaèkog hromozoma baziranog na satelitskoj
DNK (SATACs – Stable satelite DNA-based Artificial Chromosomes) definisanog
genetièkog sadraja. SATAC tehnologija se bazira na indukciji de novo formiranja
hromozoma amplifikacijom na velikoj skali, koja moe da se inicira ciljnom integracijom
strane DNK u satelit/rDNK region hromozoma domaæina. Ko-amplifikacija sekvenci
integracionog mesta ima kao rezultat de novo formiranje hromozomskih krakova i nove
hromozome koji imaju sekvence satelitske rDNK i egzogene DNK. SATACssu
heterohromatinski, ali obezbeðuju pogodnu hromozomalnu okolinu za stabilnu, trajnu
ekspresiju integrisanog genetièkog materijala.
SATACs mogu biti konstruisani, preèišæeni i preneseni u æeliju – primaoca ukljuèujuæi
oploðenu jajnu æeliju. Transgene ivotinje su uspešno kreirane preèišæenim SATACs i
utvrðena je vertikalna transmisija Zbog osnovne sliènosti elementarnih mehanizama u
æelijama eukariota, SATAC tehnologija moe da se uspešno primeni kod razlièitih vrsta,
ukljuèujuæi ljudske, animalne i biljne æelije za in vivo konstrukciju veštaèkog hromozoma
sa odabranim sekvencama.
SATELLITE DNA-BASED ARTIFICIAL CHROMOSOMES
In vivo generation of mammalian artificial chromosomes represents a reproducible and
efficient technology for construction of stable satellite DNA-based artificial chromosomes
(SATACs) with defined genetic content.
SATAC technology is based on the induction of de novo chromosome formations via
large-scale amplification, which can be initiated by targeted integration of exogenous
DNA into the satellite/rDNA region of host chromosomes. Co-amplification of sequences
of the integration site results in de novo formed chromosome arms and new
chromosomes that composed of exogenous DNA and satellite/rDNA sequences.
SATACs are heterochromatic, however, they provide a suitable chromosomal environment
for stable, persisting expression of the integrated exogenous genetic material.
SATACs can be engineered, purified and transferred into recipient cells including fertilized
eggs. Transgenic animals have successfully been generated with purified SATACs,
and the transmission of artificial chromosome through generations has been demonstrated.
Because of the basic similarities of the intrinsic cellular mechanisms between
eukaryotes, SATAC technology can successfully be applied in different species including
human, animal, and plant cells for in vivo construction of satellite DNA based artificial
chromosomes from predictable sequences.
Due to the rapid development of SATAC technology, by the development of the Artificial
Chromosome Expression System, the feasibility of the use of satellite DNA-based
artificial chromosome has been established, in different fields of gene technology. Second
generation artificial chromosomes represent a novel protein production platform
both for cellular protein production and for production of therapeutic molecules in body
fluids of transgenic animals; for development of transgenic animals with genetically
modified (humanized) tissues and organs for xenotransplantation. Also, stable SATACs
with practically unlimited carrying capacity may serve as potential vectors for animal
breeding, and for human gene therapy.