Emre TAŞKIN, Özlem KUTLU, Cüneyt KURU, Yeliz ESKİ

Düzenli Aralıklarla Bölünmüş Palindromik Tekrar Kümelerinin (CRISPR) Güncel Mikrobiyal Tanıda Kullanımı

Bulaşıcı hastalıklarda geleneksel tanı yöntemleri uzun zamandır başarıyla kullanılmaktadır. Geleneksel mikrobiyolojik tanı yöntemlerinin yanı sıra, gelişmiş doğruluk, maliyet etkinliği ve uygulama kolaylığı için yeni yöntemler geliştirilmektedir. Prokaryotik bağışıklık sistemi tipleri doğuştan gelen, adaptif ve programlı hücre ölümüdür. Son zamanlarda yeni bir bakteriyel tanı sistemi olarak CRISPR-Cas adlı system prokaryotik adaptif bağışıklık sisteminin bir parçasıdır ve yoğun olarak araştırılmaktadır. CRISPR-Cas sistemi, genetik tabanlı bakteriyel tanı yöntemlerinde sınıflandırılabilir ve aynı zamanda biyoteknolojik bir yöntem olarak da kullanılmaktadır. CRISPR-Cas sisteminin sınıflandırılması, Cas protein tipine ve hedef nükleik asit tipine (DNA veya RNA) göre yapılmaktadır. Prokaryotik hücrelerde CRISPR-Cas lokusu, tekrarlayan diziler ve spacer diziler olarak iki ana bölümden oluşur. Spacer diziler prokaryotun saldırıdan sağkalımından sonra istilacı ajanların genomundan bakteriye geçen dizilerdir. CRISPR-Cas sisteminde cas genleri prokaryotik hücrenin kendisini korumak için yabancı nükleik asidi parçalayan enzimlerdir. CRISPR-Cas sisteminin varyasyonları olarak Şu anda SHERLOCK, DETECTR ve HUDSON gibi yeni sistemler, farklı araştırma grupları tarafından geliştirilmektedir. Ayrıca CRISPR-Cas sistemi, knock-out ve knock-in sistemlerinin oluşturulmasında, transkripsiyon ve transkripsiyon sonrası seviyede gen regülasyonu gerektiren deneylerde oldukça etkilidir. CRISPR-Cas sisteminin terapötik antiviral ilaçların üretilmesinde kullanılması da planlanmaktadır. Mevcut veriler göz önüne alındığında, CRISPR-Cas, hız, düşük maliyet, yüksek doğruluk ve basit uygulama protokolü avantajları ile umut verici bir bakteriyel tanı sistemidir.

Anahtar Kelimeler:

CRISPR-Cas, Mikrobiyal tanı, Mikrobiyoloji, Biyoteknoloji

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and Its Current Applicatıons in Microbial Diagnosis

Conventional diagnostic methods have been used successfully for a long time in infectious diseases. Besides conventional microbiologic diagnosis methods, new methods are being developed for advanced accuracy, cost-effectiveness and ease of application. Prokaryotic immune system has defense types of innate, adaptive and cell suicide (programmed death). A part of prokaryotic adaptive immune system named CRISPR-Cas is under intensive research recently as a novel bacterial diagnostic system. CRISPR-Cas system can be used as a biotechnological method and can be classified in genetic based bacterial diagnostic methods. CRISPR-Cas system classification is based on included Cas protein type and the target nucleic acid type (DNA or RNA). CRISPR-Cas locus in prokaryotic cells consists of two main parts which are repeat sequences and spacer sequences. Sequences which constitutes the adaptive immunity are spacer sequences that are acquired from invading agents after survival of the prokaryote from the attack. In CRISPR-Cas system cas genes are in charge of cleaving foreign nucleic acid to defense prokaryotic cell itself. Currently new systems like SHERLOCK, DETECTR and HUDSON are developed as variations of CRISPR-Cas system by different research groups. Also, as a gene editing tool CRISPR-Cas system is highly effective in setting up knock-out and knock-in systems and in experiments which require gene regulation either in the transcriptional and post-transcriptional level. CRISPR-Cas system is planned to be used in producing therapeutic antiviral drugs. Considering current data, CRISPR-Cas is a promising bacterial diagnostic system with all its advantages of rapidity, lower cost, accuracy and simple application protocol.

Keywords:

CRISPR-Cas, Bacterial diagnosis, microbiology, biotechnology,

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