誘導CRISPRスペーサー取得による分子記録

Seth L Shipman¹, Jeff Nivala², Jeffrey D Macklis³

¹Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA. Department of Stem Cell and Regenerative Biology, Center for Brain Science, and Harvard Stem Cell Institute, Harvard University, Bauer Laboratory 103, Cambridge, MA 02138, USA. Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA.

Science (New York, N.Y.)

|June 11, 2016

まとめ

科学者たちは E. coli のCRISPR-Casシステムを設計し分子現象を DNA 配列として記録しましたこれは生物学や合成装置の応用のために安定した細胞の歴史を生み出します

科学分野:

微生物学
分子生物学
合成生物学

背景:

細胞内の分子イベントの安定した記録は細胞の歴史を理解するために不可欠です
CRISPR-Casシステムは標的型DNAの操作と記録の可能性を秘めています

研究の目的:

合成DNAの配列を記録するためのE大腸菌のタイプI-E CRISPR-Casシステムを設計する.
CRISPR-Casの適応を基にマルチモダル細胞内記録装置を開発する.

主な方法:

E. coliのタイプI-E CRISPR-Casシステムを活用して,合成DNAの取得を介した.
Cas1-Cas2複合体によるプロトスペーサーの隣接モチーフ認識を修正するために,適用された指向進化.
2つの異なるモードで同時に録音できます.

主要な成果:

細菌のゲノム内の特定のDNA配列の安定した記録を生成する能力を示した.
プロトスペーサーの隣接モチーフ認識を成功裏に改変し記録能力を強化しました
CRISPR-Casの適応に不可欠なスペースアキシビションプロセスに関する新しい洞察を明らかにしました

結論:

設計されたCRISPR-Casシステムは多様性細胞内記録装置の基盤として機能します
この技術は,発達生物学と合成装置における潜在的な応用を持つ長い細胞の歴史の調査を可能にします.
この発見はCRISPR-Casの適応メカニズムの理解を深めるものです

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