Gen terapi yöntemleri: Fiziksel ve kimyasal metodlar

Öz

Gen terapisi, genetik temeli bulunan hastalıkların tedavisinde stratejiler geliştirmek için kullanılan ve günümüzde tedavisi olmayan hastalıklar için umut vaat eden bir yöntemdir. Başarılı bir gen terapisi, ilgili transgenleri içeren plazmidlerin hedeflenmiş hücrelere transfeksiyonunu gerektirir. Gen tedavisi çalışmalarında karşılaşılan en önemli sorun olan DNA moleküllerinin hedef hücrelere ulaştırılması, bu alanda çalışan tüm araştırmacıları etkili bir yol bulmaya yöneltmiştir. DNA’nın hücrelere girme yeteneğinin kısıtlı oluşu ve DNA’nın enzimatik degredasyona uğrama ihtimali nedeniyle DNA transfeksiyonu çoğunlukla bir vektör aracılığıyla gerçekleştirilir. Bunlar, viral vektörler ve viral olmayan vektörler olarak iki gruba ayrılmaktadır. Adenovirüs, adeno-assosiye virüs, herpes simpleks ve retrovirüsler viral vektörlerin başta gelen örnekleridir. Viral olmayan vektörler ise kendi içinde fiziksel ve kimyasal olarak ayrılırlar. Fiziksel metotlar; mikroenjeksiyon, partikül bombardımanı gen tabancası, elektroporasyon, sonoporasyon, laser ışıması ve magnetofeksiyondur. Kimyasal metotlar ise viral vektörlere alternatif olarak ortaya çıkmış olan lipozomları kapsarlar. Bu vektörler, hücre çekirdeğine yapılacak gen transferini arttırmak amacıyla üç önemli özelliğe sahip olmalıdırlar. Bunlar DNA’nın negatif yükünü maskelemek, DNA molekülünü sıkıştırarak taşıyacağı kargoyu kompakt hale getirmek ve onu hücre içi nükleaz degredasyonundan korumaktır. Lipozomlar gibi viral olmayan transfeksiyon sistemleri virüslerden daha fazla tercih edilir. Çünkü bunlar immünojenik değildir, yapımı kolaydır ve endüstriyel üretim amacıyla ölçek büyütme işlemi daha basittir. Lipozomal taşıma araçları morfoloji ve salınım karakteristiği açısından çeşitlilik sağlar; doku hedeflemede kullanılabilir ve plazmid DNA’yı degredatif nükleazların saldırılarından koruyabilir. 1987’de potansiyel taşıma sistemi olarak tanımlanmalarından beri, DNA-katyonik lipit kompleksleri çeşitli hücre tiplerinde farklı DNA aktarım protokollerinde kullanılmıştır ve halen gen terapisinin klinik çalışmaları için araştırılmaktadır. Bu derleme, yeni ve gelecek vaat eden bir teknik olarak kanser ve genetik temelli hastalıkların tedavisinde kullanılması hedeflenen gen terapisindeki kimyasal ve fiziksel yöntemleri anlatmaktadır

Anahtar Kelimeler

• gen terapisi, lipozomal vektörler, plazmid DNA, gen transferi

Gene therapy techniques: Physical and chemical methods

Abstract

Gene therapy is used for developing strategies for the treatment of genetic diseases and it is a promising technique for people with incurable diseases. A successful gene therapy includes the transfection of plasmids with related transgenes into the target cells. The transfer of the DNA molecule to target cell is the main problem of the gene therapy studies which directed researchers to find an effective way of transfection. The transfection of DNA is commonly achieved by a vector because of the limited insertion ability of the DNA into the cells and the possibility of enzymatic degradation of DNA molecule. These vectors are grouped into two categories as viral and non-viral. Adenovirus, adenoassociated virus, herpes simplex and retrovirus are the main examples of viral vectors. Non-viral vectors are grouped into two as physical and chemical methods. The physical methods are microinjection, particle bombardment - gene gun, electroporation, sonoporation, laser beam and magnetofection. The chemical methods are consisted of liposomes which developed as an alternative to the viral vectors. These vectors must have three important features for the transfer of the related gene into the cell nucleus. Those are disguising the negative charge of the DNA, condensing the DNA molecule and protecting it from the intracellular nuclease activity. içi nükleaz degredasyonundan korumaktır. Lipozomlar gibi viral olmayan transfeksiyon sistemleri virüslerden daha fazla tercih edilir. Çünkü bunlar immünojenik değildir, yapımı kolaydır ve endüstriyel üretim amacıyla ölçek büyütme işlemi daha basittir. Lipozomal taşıma araçları morfoloji ve salınım karakteristiği açısından çeşitlilik sağlar; doku hedeflemede kullanılabilir ve plazmid DNA’yı degredatif nükleazların saldırılarından koruyabilir. 1987’de potansiyel taşıma sistemi olarak tanımlanmalarından beri, DNA-katyonik lipit kompleksleri çeşitli hücre tiplerinde farklı DNA aktarım protokollerinde kullanılmıştır ve halen gen terapisinin klinik çalışmaları için araştırılmaktadır. Bu derleme, yeni ve gelecek vaat eden bir teknik olarak kanser ve genetik temelli hastalıkların tedavisinde kullanılması hedeflenen gen terapisindeki kimyasal ve fiziksel yöntemleri anlatmaktadır

Keywords

• gene therapy, liposomal vectors, plasmid DNA, gene transfer

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