Related Experiment Video
Updated: Aug 15, 2026
High-Throughput Robotically Assisted Isolation of Temperature-sensitive Lethal Mutants in Chlamydomonas reinhardtii
Published on: December 5, 2016
Temperature-sensitive cell-lethal mutants of drosophila: isolation and characterization
Abstract:
One hundred and twenty-one sensitive (ts) sex-linked lethals were screened by means of X-ray-induced somatic crossing over to determine if any were ts cell-lethal mutants. Cell-lethal mutations were identified by their ability to block the development of homozygous clones when raised under restrictive conditions (29degrees). Twenty-two ts cell-lethal mutants were isolated and categorized into three classes, depending upon the patterns of damage observed in larval and imaginal tissues. The phenotypes produced by these mutations ranged from those which affected only a limited set of structures (i.e., genital discs only) to those which affected diverse tissues at all stages of the life cycle. Each mutation has its own characteristic time-dependent pattern, frequency, and type of damage. All the mutations affect imaginal tissue, but only one-third of the mutations affect both larval and imaginal tissue. The fastest-acting lethals need 15 hours at the restrictive temperature to kill the cells and the slowest-acting lethals require at least 48 hours. By choosing the appropriate mutant and by manipulating the times of exposure to the restrictive temperature, it has proven possible to produce duplications and deficiencies in specific structures of the adult. A mechanism by which lethality might yield such structures is suggested. In addition, 15 of the mutants are ts female sterile mutants. Only one of these 15 mutants can recover its fertility when shifted back down to the permissive temperature (22degrees).
Insights
Researchers screened temperature-sensitive (ts) sex-linked lethals for cell-lethal mutations using X-ray-induced somatic crossing over. Twenty-two ts cell-lethal mutants were identified, impacting larval and imaginal tissues with varying damage patterns and timelines.
Area of Science:
- Developmental Biology
- Genetics
- Molecular Biology
Background:
- Temperature-sensitive (ts) mutations are valuable tools for studying gene function.
- Somatic crossing over is a technique used to uncover recessive lethal mutations.
Purpose of the Study:
- To identify and characterize temperature-sensitive (ts) cell-lethal mutations.
- To investigate the developmental effects of these mutations on larval and imaginal tissues.
- To explore the potential for manipulating these mutations to create specific adult structures.
Main Methods:
- Screening of 121 ts sex-linked lethals using X-ray-induced somatic crossing over.
- Identification of cell-lethal mutants by their ability to block homozygous clone development at restrictive temperature (29°C).
- Categorization of 22 isolated ts cell-lethal mutants based on observed damage patterns in larval and imaginal tissues.
Main Results:
- Twenty-two ts cell-lethal mutants were identified, falling into three distinct classes based on tissue damage.
- Mutations exhibited diverse phenotypes, affecting specific structures or multiple tissues across developmental stages.
- Lethality onset varied from 15 to over 48 hours at the restrictive temperature.
- Fifteen mutants were also identified as ts female sterile, with only one recovering fertility upon return to permissive temperature (22°C).
Conclusions:
- The study successfully identified and characterized a set of ts cell-lethal mutants with distinct developmental impacts.
- The findings suggest that specific mutations and temperature manipulations can be used to generate structural duplications and deficiencies in adult organisms.
- The identified ts female sterile mutants offer further avenues for research into fertility regulation.
