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Related Experiment Video

Updated: May 31, 2026

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
08:37

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FLInt 2.0: Robust and customizable single shot integration in C. elegans.

Nawaphat Malaiwong1, Porhathai Malaiwong1, Chloe Kim1

  • 1Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06511, USA.

G3 (Bethesda, Md.)
|May 28, 2026
PubMed
Summary

FLInt 2.0 enhances transgene integration in C. elegans by reducing false positives. This refined CRISPR-Cas9 method improves screening efficiency for generating stable transgenic lines.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Developmental Biology

Background:

  • Transgenesis in *C. elegans* allows precise gene expression control.
  • Fluorescent Landmark Interference (FLInt) facilitates transgene integration via CRISPR-Cas9.
  • FLInt 1.0 suffered from high false-positive rates, complicating screening.

Purpose of the Study:

  • To develop an improved FLInt strategy (FLInt 2.0) to minimize false positives in transgene integration.
  • To enhance the efficiency and reliability of generating stable transgenic *C. elegans* lines.

Main Methods:

  • FLInt 2.0 utilizes targeted CRISPR-Cas9 cutting of fluorescent protein landing sites.
  • Non-fluorescent F2 animals are used to identify stable, multi-copy transgenic lines.
  • Investigated the impact of DNA structure (linear vs. circular) on integration efficiency.
Keywords:
C. elegansCRISPRWormBasemultiplex gene expressionsafe landing sitestransgenesis

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Last Updated: May 31, 2026

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
08:37

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

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High-throughput Screening and Biosensing with Fluorescent C. elegans Strains

Published on: May 19, 2011

A Semi-high-throughput Imaging Method and Data Visualization Toolkit to Analyze C. elegans Embryonic Development
06:49

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Main Results:

  • FLInt 2.0 significantly reduces false positives compared to FLInt 1.0.
  • Non-fluorescent F2 generation reliably indicates successful transgene integration.
  • Linear DNA substrates enhance transgene array formation and insertion efficiency.
  • Subsequent Cas9 targeting allows fine-tuning of expression levels in multicopy lines.

Conclusions:

  • FLInt 2.0 offers a robust, visually guided method for site-specific transgene integration in *C. elegans*.
  • This approach streamlines the generation of transgenic lines, increasing experimental throughput.
  • The strategy provides a generalizable framework for improving transgene integration techniques.