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CRISPR01:59

CRISPR

57.8K
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...
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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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...
19.1K
CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

1.8K
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
1.8K
RNA Interference01:23

RNA Interference

28.0K
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...
28.0K
Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

14.8K
Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
14.8K
RNA Structure01:23

RNA Structure

79.1K
Overview
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
79.1K

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Related Experiment Video

Updated: Jan 30, 2026

Generation of Brown Fat-Specific Knockout Mice Using a Combined Cre-LoxP, CRISPR-Cas9, and Adeno-Associated Virus Single-Guide RNA System
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Generation of Brown Fat-Specific Knockout Mice Using a Combined Cre-LoxP, CRISPR-Cas9, and Adeno-Associated Virus Single-Guide RNA System

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Rapid Generation of Long Noncoding RNA Knockout Mice Using CRISPR/Cas9 Technology.

Nils R Hansmeier1,2, Pia J M Widdershooven3, Sajjad Khani4,5,6

  • 1Max Planck Institute for Metabolism Research, Gleueler Strasse 50, 50931 Cologne, Germany. Nils.Hansmeier@sf.mpg.de.

Non-Coding RNA
|January 26, 2019
PubMed
Summary

We developed a CRISPR/Cas9 method to create long noncoding RNA (lncRNA) knockout mice. This efficient process allows for rapid generation and validation of lncRNA loss-of-function models for studying gene regulation.

Keywords:
long noncoding RNA, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated genome engineering, knockout mice

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Area of Science:

  • Molecular Biology
  • Genetics
  • Genomics

Background:

  • Long noncoding RNAs (lncRNAs) are key regulators of gene expression in development and disease.
  • Limited availability of lncRNA loss-of-function models hinders in vivo studies of their physiological roles.
  • CRISPR/Cas9 technology enables targeted genome editing but requires specific strategies for lncRNA targeting.

Purpose of the Study:

  • To establish a robust, step-wise protocol for generating and validating lncRNA loss-of-function mouse models using CRISPR/Cas9.
  • To demonstrate the utility of this method by creating a knockout mouse model for the liver-enriched lncRNA Gm15441.
  • To provide guidelines for lncRNA target selection and validation procedures.

Main Methods:

  • Utilized CRISPR/Cas9 genome engineering for targeted disruption of lncRNA loci.
  • Developed specific strategies for targeting lncRNAs, considering their lack of open-reading frames.
  • Performed in vitro validation of single guide RNAs (sgRNAs) and assessed in vivo gene-targeting efficiency and knockout confirmation.

Main Results:

  • Successfully generated a mouse model deficient for the liver-enriched lncRNA Gm15441.
  • Gm15441 was found to be downregulated in metabolic disease and responsive to feeding/fasting transitions.
  • The entire process from target selection to validation was completed within 18-20 weeks with minimal hands-on time (<10 days).

Conclusions:

  • The described CRISPR/Cas9-based protocol provides an efficient and effective approach for generating lncRNA loss-of-function mouse models.
  • This methodology facilitates in-depth investigation of lncRNA functions in organismal homeostasis and pathophysiology.
  • The rapid generation of these models accelerates research into the roles of lncRNAs in development and disease.