Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Experimental RNAi02:15

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...
Genetic Screens02:46

Genetic Screens

Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Applications of genome editing technologies in the treatment of human diseases.

Gene therapy·2026
Same author

Molecular mechanism of <i>miR-27a-3p</i> targeting <i>FBXW7</i> regulating the malignant behavior of osteosarcoma cells.

Translational cancer research·2026
Same author

Transcription and cohesin direct domain boundary spatial positioning and are linked to Friedreich's ataxia.

Molecular cell·2026
Same author

Abscisic acid modulates genotype-specific root cell-wall cadmium retention in pepper: evidence from adsorption kinetics and transcriptomic networks.

Plant physiology and biochemistry : PPB·2026
Same author

Exosomes Derived From Mesenchymal Stem Cells Regulating Myotube Cell Atrophy via NF-κB Signaling Pathway.

Stem cells international·2026
Same author

Artificial intelligence in immunotherapy: revolutionizing diagnostic and therapeutic applications in cancer and autoimmune diseases.

Clinical and experimental medicine·2026

Related Experiment Video

Updated: Jul 1, 2026

MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells
09:12

MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells

Published on: December 21, 2011

A random shRNA-encoding library for phenotypic selection and hit-optimization.

Yongping Wang1, Yun E Wang, M Grazia Cotticelli

  • 1Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.

Plos One
|September 10, 2008
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel, completely random RNA interference (RNAi) library. This tool identified sequences enhancing cell survival under stress, enabling unbiased discovery of gene silencing functions for therapeutics.

More Related Videos

Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype
14:51

Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype

Published on: June 17, 2022

Primer-Free Aptamer Selection Using A Random DNA Library
11:14

Primer-Free Aptamer Selection Using A Random DNA Library

Published on: July 26, 2010

Related Experiment Videos

Last Updated: Jul 1, 2026

MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells
09:12

MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells

Published on: December 21, 2011

Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype
14:51

Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype

Published on: June 17, 2022

Primer-Free Aptamer Selection Using A Random DNA Library
11:14

Primer-Free Aptamer Selection Using A Random DNA Library

Published on: July 26, 2010

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • RNA interference (RNAi) typically targets single genes via canonical pathways.
  • Endogenous microRNAs (miRNAs) often target multiple genes and can use non-canonical pathways, including gene activation.
  • Existing RNAi libraries are limited in scope compared to natural miRNA functions.

Purpose of the Study:

  • To design and validate the first completely random nucleotide-level shRNA-encoding library.
  • To identify novel shRNA sequences with desired phenotypic effects.
  • To enable unbiased screening for gene silencing functions beyond canonical pathways.

Main Methods:

  • Designed and synthesized a completely random shRNA-encoding library.
  • Screened the library in an IL3-dependent cell line (FL5.12) under IL3 withdrawal conditions.
  • Employed random mutagenesis and re-screening for hit optimization under stringent conditions.

Main Results:

  • Identified shRNA sequences that doubled cell survival upon IL3 withdrawal.
  • Optimized a lead sequence through random mutagenesis, enhancing its potency.
  • Demonstrated that a specific nucleotide change and stem mismatch improved shRNA efficacy.

Conclusions:

  • The random shRNA library enables unbiased selection and optimization of sequences conferring specific phenotypes.
  • This approach facilitates the discovery of novel gene silencing functions.
  • The library holds potential for developing therapeutics and biological tools.