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CRISPR

CRISPR

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...

siRNA - Small Interfering RNAs

siRNA - Small Interfering RNAs

Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...

Experimental RNAi

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...

CRISPR/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

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...

RNA Interference

RNA Interference

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...

lncRNA - Long Non-coding RNAs

lncRNA - Long Non-coding RNAs

In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA...

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Related Experiment Video

Updated: Nov 1, 2025

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells

Published on: May 30, 2025

CRISPR/Cas9 to Silence Long Non-Coding RNAs.

Ingrid Arctander Rosenlund¹, George A Calin^2,3, Mihnea P Dragomir⁴

¹Department of Medical Biology, Faculty of Health Sciences, UiT-The Arctic University of Norway, Tromsø, Norway.

Methods in Molecular Biology (Clifton, N.J.)

|June 23, 2021

Summary

This study details CRISPR/Cas9 gene editing to knock out long non-coding RNAs (lncRNAs). The method uses electroporation for efficient in vitro functional characterization of these poorly understood transcripts.

Keywords:

CRISPR/Cas9 Electroporation Gene editing Knockout Long non-coding RNA

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Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells

Published on: September 1, 2019

Overexpressing Long Noncoding RNAs Using Gene-activating CRISPR

Overexpressing Long Noncoding RNAs Using Gene-activating CRISPR

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Last Updated: Nov 1, 2025

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells

Published on: May 30, 2025

Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells

Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells

Published on: September 1, 2019

Overexpressing Long Noncoding RNAs Using Gene-activating CRISPR

Overexpressing Long Noncoding RNAs Using Gene-activating CRISPR

Published on: March 1, 2019

Area of Science:

Molecular Biology
Genetics
RNA Biology

Background:

Long non-coding RNAs (lncRNAs) are a poorly characterized class of transcripts.
Functional characterization of lncRNAs is crucial for understanding their roles.
CRISPR/Cas gene editing offers a powerful tool for genetic manipulation.

Purpose of the Study:

To describe a methodology for the CRISPR/Cas9-mediated knockout (KO) of lncRNAs.
To establish an efficient in vitro system for lncRNA functional studies.
To address the challenges of lncRNA KO due to incomplete annotation.

Main Methods:

Utilizing CRISPR/Cas9 gene editing technology for lncRNA knockout.
Employing single guide RNAs (sgRNAs) and Cas9 enzyme for targeted gene disruption.
Transfecting cells with presynthesized sgRNAs and Cas9 via electroporation for in vitro application.

Main Results:

Demonstrated a methodology for complete lncRNA knockout.
Successfully applied CRISPR/Cas9 KO in an in vitro setting.
Established electroporation as an efficient transfection method for this application.

Conclusions:

CRISPR/Cas9 gene editing is a viable and efficient strategy for lncRNA knockout.
The described methodology facilitates functional characterization of lncRNAs.
This approach aids in the study of lncRNA roles in biological processes.