RNAi in C. elegans: Soaking in the Genome sequence

Science

Volumn 282, Issue 5388, 1998, Pages 430-431

  Tabara, Hiroaki ^a   Grishok, Alla ^a   Mello, Craig C ^a  

^a UNIVERSITY OF MASSACHUSETTS MEDICAL SCHOOL   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

COMPLEMENTARY RNA; DNA; DOUBLE STRANDED RNA; MESSENGER RNA; RNA;

CAENORHABDITIS ELEGANS; ESCHERICHIA COLI; GENE REPRESSION; GENOME; NONHUMAN; PRIORITY JOURNAL; SHORT SURVEY;

ANIMALS; CAENORHABDITIS ELEGANS; GENE EXPRESSION REGULATION; GENES, HELMINTH; MUTATION; PHENOTYPE; RNA, ANTISENSE; RNA, DOUBLE-STRANDED; RNA, HELMINTH; RNA, MESSENGER; TRANSCRIPTION, GENETIC;

CAENORHABDITIS ELEGANS; ESCHERICHIA COLI; INVERTEBRATA;

EID: 0032538391     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.282.5388.430     Document Type: Short Survey

References (17)

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5. A. Grishok and C. C. Mello, unpublished results. We do not know whether this long-term transmission, which is always somewhat less penetrant than the interference observed in the first two generations, involves the same or a secondary interference mechanism.
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12. C. C. Mello, unpublished observations.
13. In one case, we found that two genes that were 80% identical showed clear evidence of cross-interference. Specific interference was achieved by diluting the injected RNA from 1 mg/ml to 100 μg/ml. (12).
14. L4-stage hermaphrodites were washed in 0.2 M sucrose and 0.1× phosphate-buffered saline, and transferred into 10 μl of the same buffer in a siliconized tube. In another siliconized tube, 4 μl of dsRNA (3.8 mg/ml) and 1 μl liposome (Lipofectin, Gibco-BRL) were vigorously mixed. Fifteen worms were added to the RNA-liposome mixture, resulting in a total volume of 15 μl and a final RNA concentration of 1 mg/ml. After 24 hours, the worms were transferred to an agar plate with E. coli and cultured until midadulthood. Similar levels of interference were obtained by feeding worms F. coli expressing both strands of pos-1 RNA (10).
15. H. Tabara and C. C. Mello, unpublished observations.
16. M. K. Montgomery and A. Fire, Trends Genet. 14, 255 (1998).
17. We thank J. Bosher, M. Labouesse, L. Timmons and A. Fire for communicating unpublished results. Work in this laboratory is supported by NIH grant R01 HD33769-01, a pilot and feasibility project grant DK32520-15, and a Pew scholarship to C.C.M.