関連する実験動画
Updated: Jan 23, 2026

06:48
CRISPR Guide RNA Cloning for Mammalian Systems
Published on: October 2, 2018
72.9K
CRISPR関連トランポザースによるRNA誘導DNA挿入
Jonathan Strecker1,2,3,4, Alim Ladha1,2,3,4, Zachary Gardner1,2,3,4
1Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
まとめ
研究者たちは新しいCRISPR-Casシステム (ShCAST) を発見し 細菌のゲノムにDNAを正確に挿入しました このRNA誘導のDNA転置システムは,ホスト細胞の修復を必要とせずに,高い効率を達成し,遺伝子編集技術を進歩させています.
科学分野:
- 分子生物学
- 遺伝学
- 微生物学
背景:
- CRISPR-Cas核酸は,核酸の操作に不可欠である.
- CRISPR技術を用いた標的型DNA挿入は,宿主細胞の修復メカニズムに依存しているため,課題に直面しています.
研究 の 目的:
- サイアノバクテリア (ShCAST) の新型CRISPR関連トランポゼの特徴を特定する.
- 細菌のゲノムにRNA誘導のDNA転移と挿入のためのShCASTの能力を評価する.
主な方法:
- Tn7型のトランポザースサブユニットとCas12kエフェクターを含むShCASTシステムの特徴.
- ShCASTのDNA挿入効率とEscherichia coliにおけるサイト特異性を評価する.
- RNAによるDNA転移のメカニズムを調査する.
主要な成果:
- ShCASTはRNA誘導のDNAトランスポジションを触媒化し,DNAセグメント60~66の塩基対をプロトスペーサーの下流に挿入します.
- ShCASTは,陽性選択なしに,E. coliのゲノムに最大80%の統合効率を達成しました.
- ShCASTによる単方向のDNA挿入が実証された.
結論:
- ShCASTはトランスポザース活性を持つ新しいCRISPR- Casシステムです.
- このシステムにより 精密で効率的な 選択独立のDNA挿入が可能になります
- CRISPR-Casの機能的多様性についての理解を広げ,精密ゲノム工学の新しいパラダイムを提供します.
関連する概念動画
CRISPR
57.6K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
57.6K
CRISPR and crRNAs
18.8K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
18.8K
DNA-only Transposons
17.3K
DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
The donor site from where the transposon is excised is either degraded or...
17.3K
Eukaryotic RNA Polymerases
26.8K
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
All three eukaryotic RNAPs require specific transcription factors, of which the...
26.8K
DNA Replication
58.9K
DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied. After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
Replication in Prokaryotes
DNA replication...
Replication in Prokaryotes
DNA replication...
58.9K
RNA Interference
27.9K
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
27.9K

