Detection of CRISPR adaptation

Anna Shiriaeva, Ivan Fedorov, Danylo Vyhovskyi, Konstantin Severinov

    Research output: Contribution to journalReview articlepeer-review

    7 Citations (Scopus)

    Abstract

    Prokaryotic adaptive immunity is built when short DNA fragments called spacers are acquired into CRISPR (clustered regularly interspaced short palindromic repeats) arrays. CRISPR adaptation is a multistep process which comprises selection, generation, and incorporation of prespacers into arrays. Once adapted, spacers provide immunity through the recognition of complementary nucleic acid sequences, channeling them for destruction. To prevent deleterious autoimmunity, CRISPR adaptation must therefore be a highly regulated and infrequent process, at least in the absence of genetic invaders. Over the years, ingenious methods to study CRISPR adaptation have been developed. In this paper, we discuss and compare methods that detect CRISPR adaptation and its intermediates in vivo and propose suppressing PCR as a simple modification of a popular assay to monitor spacer acquisition with increased sensitivity.

    Original languageEnglish
    Pages (from-to)257-269
    Number of pages13
    JournalBiochemical Society Transactions
    Volume48
    Issue number1
    DOIs
    Publication statusPublished - 2020

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