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The C. Elegans Heterochronic Gene Lin-46 Affects Developmental Timing At Two Larval Stages And Encodes A Relative Of The Scaffolding Protein Gephyrin., Anita Pepper, Jill Mccane, Kevin Kemper, Denise Yeung, Rosalind Lee, Victor Ambros, Eric Moss Apr 2004

The C. Elegans Heterochronic Gene Lin-46 Affects Developmental Timing At Two Larval Stages And Encodes A Relative Of The Scaffolding Protein Gephyrin., Anita Pepper, Jill Mccane, Kevin Kemper, Denise Yeung, Rosalind Lee, Victor Ambros, Eric Moss

Victor R. Ambros

The succession of developmental events in the C. elegans larva is governed by the heterochronic genes. When mutated, these genes cause either precocious or retarded developmental phenotypes, in which stage-specific patterns of cell division and differentiation are either skipped or reiterated, respectively. We identified a new heterochronic gene, lin-46, from mutations that suppress the precocious phenotypes caused by mutations in the heterochronic genes lin-14 and lin-28. lin-46 mutants on their own display retarded phenotypes in which cell division patterns are reiterated and differentiation is prevented in certain cell lineages. Our analysis indicates that lin-46 acts at a step immediately downstream …


Expression Profiling Of Mammalian Micrornas Uncovers A Subset Of Brain-Expressed Micrornas With Possible Roles In Murine And Human Neuronal Differentiation., Lorenzo F. Sempere, Sarah Freemantle, Ian Pitha-Rowe, Eric Moss, Ethan Dmitrovsky, Victor R. Ambros Feb 2004

Expression Profiling Of Mammalian Micrornas Uncovers A Subset Of Brain-Expressed Micrornas With Possible Roles In Murine And Human Neuronal Differentiation., Lorenzo F. Sempere, Sarah Freemantle, Ian Pitha-Rowe, Eric Moss, Ethan Dmitrovsky, Victor R. Ambros

Victor R. Ambros

Background The microRNAs (miRNAs) are an extensive class of small noncoding RNAs (18 to 25 nucleotides) with probable roles in the regulation of gene expression. In Caenorhabditis elegans, lin-4 and let-7 miRNAs control the timing of fate specification of neuronal and hypodermal cells during larval development. lin-4, let-7 and other miRNA genes are conserved in mammals, and their potential functions in mammalian development are under active study. Results In order to identify mammalian miRNAs that might function in development, we characterized the expression of 119 previously reported miRNAs in adult organs from mouse and human using northern blot analysis. Of …


A Short History Of A Short Rna, Victor Ambros, Rosalind Lee, Rhonda Feinbaum Jan 2004

A Short History Of A Short Rna, Victor Ambros, Rosalind Lee, Rhonda Feinbaum

Victor R. Ambros

Comment on: The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. [Cell. 1993]


Identification Of Micrornas And Other Tiny Noncoding Rnas By Cdna Cloning, Victor Ambros, Rosalind Lee Dec 2003

Identification Of Micrornas And Other Tiny Noncoding Rnas By Cdna Cloning, Victor Ambros, Rosalind Lee

Victor R. Ambros

MicroRNAs (miRNAs) and other small RNAs can be identified by cloning and sequencing cDNAs prepared from the ∼22-nt fraction of total RNA. Methods are described for the construction of cDNA libraries from small noncoding RNAs through the use of T4 RNA ligase, reverse transcriptase, and polymerase chain reaction. cDNAs are cloned in λ or plasmid vectors, and the sequences are compared to annotated genomic sequence databases, and analyzed by RNA folding programs to distinguish miRNA sequences from other small RNAs of similar size. Northern blot hybridization is used to confirm the expression of small RNAs in vivo.


Role Of Micrornas In Plant And Animal Development, Victor Ambros, James Carrington Jul 2003

Role Of Micrornas In Plant And Animal Development, Victor Ambros, James Carrington

Victor R. Ambros

Small RNAs, including microRNAs (miRNAs) and short interfering RNAs (siRNAs), are key components of an evolutionarily conserved system of RNA-based gene regulation in eukaryotes. They are involved in many molecular interactions, including defense against viruses and regulation of gene expression during development. miRNAs interfere with expression of messenger RNAs encoding factors that control developmental timing, stem cell maintenance, and other developmental and physiological processes in plants and animals. miRNAs are negative regulators that function as specificity determinants, or guides, within complexes that inhibit protein synthesis (animals) or promote degradation (plants) of mRNA targets.


Microrna Pathways In Flies And Worms: Growth, Death, Fat, Stress, And Timing, Victor Ambros Jun 2003

Microrna Pathways In Flies And Worms: Growth, Death, Fat, Stress, And Timing, Victor Ambros

Victor R. Ambros

Drosophila geneticists have uncovered roles for microRNAs in the coordination of cell proliferation and cell death during development, and in stress resistance and fat metabolism. In C. elegans, a homolog of the well-known fly developmental regulator hunchback acts downstream of the microRNAs lin-4 and let-7 in a pathway controlling developmental timing.


Micrornas And Other Tiny Endogenous Rnas In C. Elegans, Victor Ambros, Rosalind Lee, Ann Lavanway, Peter Williams, David Jewell May 2003

Micrornas And Other Tiny Endogenous Rnas In C. Elegans, Victor Ambros, Rosalind Lee, Ann Lavanway, Peter Williams, David Jewell

Victor R. Ambros

BACKGROUND: MicroRNAs (miRNAs) are small noncoding RNAs that are processed from hairpin precursor transcripts by Dicer. miRNAs probably inhibit translation of mRNAs via imprecise antisense base-pairing. Small interfering RNAs (siRNAs) are similar in size to miRNAs, but they recognize targets by precise complementarity and elicit RNA-mediated interference (RNAi). We employed cDNA sequencing and comparative genomics to identify additional C. elegans small RNAs with properties similar to miRNAs and siRNAs. RESULTS: We found three broad classes of small RNAs in C. elegans: (1) 21 new miRNA genes (we estimate that C. elegans contains approximately 100 distinct miRNA genes, about 30% of …


A Uniform System For Microrna Annotation, Victor R. Ambros, Bonnie Bartel, David P. Bartel, Christopher B. Burge, James C. Carrington, Xuemei Chen, Gideon Dreyfuss, Sean R. Eddy, Sam Griffiths-Jones, Mhairi Marshall, Marjori Matzke, Gary Ruvkun, Thomas Tuschl Feb 2003

A Uniform System For Microrna Annotation, Victor R. Ambros, Bonnie Bartel, David P. Bartel, Christopher B. Burge, James C. Carrington, Xuemei Chen, Gideon Dreyfuss, Sean R. Eddy, Sam Griffiths-Jones, Mhairi Marshall, Marjori Matzke, Gary Ruvkun, Thomas Tuschl

Victor R. Ambros

MicroRNAs (miRNAs) are small noncoding RNA gene products about 22 nt long that are processed by Dicer from precursors with a characteristic hairpin secondary structure. Guidelines are presented for the identification and annotation of new miRNAs from diverse organisms, particularly so that miRNAs can be reliably distinguished from other RNAs such as small interfering RNAs. We describe specific criteria for the experimental verification of miRNAs, and conventions for naming miRNAs and miRNA genes. Finally, an online clearinghouse for miRNA gene name assignments is provided by the Rfam database of RNA families.


The Expression Of The Let-7 Small Regulatory Rna Is Controlled By Ecdysone During Metamorphosis In Drosophila Melanogaster, Lorenzo Sempere, Edward Dubrovsky, Veronica Dubrovskaya, Edward Berger, Victor Ambros Mar 2002

The Expression Of The Let-7 Small Regulatory Rna Is Controlled By Ecdysone During Metamorphosis In Drosophila Melanogaster, Lorenzo Sempere, Edward Dubrovsky, Veronica Dubrovskaya, Edward Berger, Victor Ambros

Victor R. Ambros

In Caenorhabditis elegans, the heterochronic pathway controls the timing of developmental events during the larval stages. A component of this pathway, the let-7 small regulatory RNA, is expressed at the late stages of development and promotes the transition from larval to adult (L/A) stages. The stage-specificity of let-7 expression, which is crucial for the proper timing of the worm L/A transition, is conserved in Drosophila melanogaster and other invertebrates. In Drosophila, pulses of the steroid hormone 20-hydroxyecdysone (ecdysone) control the timing of the transition from larval to pupal to adult stages. To test whether let-7 expression is regulated by ecdysone …


Micrornas: Tiny Regulators With Great Potential, Victor Ambros Dec 2001

Micrornas: Tiny Regulators With Great Potential, Victor Ambros

Victor R. Ambros

Animal genomes contain an abundance of small genes that produce regulatory RNAs of about 22 nucleotides in length. These microRNAs are diverse in sequence and expression patterns, and are evolutionarily widespread, suggesting that they may participate in a wide range of genetic regulatory pathways.


An Extensive Class Of Small Rnas In Caenorhabditis Elegans, Rosalind Lee, Victor Ambros Oct 2001

An Extensive Class Of Small Rnas In Caenorhabditis Elegans, Rosalind Lee, Victor Ambros

Victor R. Ambros

The lin-4 and let-7 antisense RNAs are temporal regulators that control the timing of developmental events in Caenorhabditis elegans by inhibiting translation of target mRNAs. let-7 RNA is conserved among bilaterian animals, suggesting that this class of small RNAs [microRNAs (miRNAs)] is evolutionarily ancient. Using bioinformatics and cDNA cloning, we found 15 new miRNA genes in C. elegans. Several of these genes express small transcripts that vary in abundance during C. elegans larval development, and three of them have apparent homologs in mammals and/or insects. Small noncoding RNAs of the miRNA class appear to be numerous and diverse.


Development. Dicing Up Rnas, Victor Ambros Aug 2001

Development. Dicing Up Rnas, Victor Ambros

Victor R. Ambros

Comment on: A cellular function for the RNA-interference enzyme Dicer in the maturation of the let-7 small temporal RNA. [Science. 2001]


The Temporal Control Of Cell Cycle And Cell Fate In Caenorhabditis Elegans, Victor Ambros Dec 2000

The Temporal Control Of Cell Cycle And Cell Fate In Caenorhabditis Elegans, Victor Ambros

Victor R. Ambros

The nematode Caenorhabditis elegans develops through two major phases: the first phase, embryogenesis, consists of a rapid series of cleavage cell divisions leading to morphogenesis of a first stage larva. The second phase is postembryonic development, which consists of developmentally regulated cell cycles that occur during the four larval stages leading to the adult. Precursor cells set aside during embryogenesis divide through stereotypical cell lineage patterns during the four larval stages to generate larval and adult structures. The precise timing of the postembryonic cell divisions is under strict control, in most cases with a developmentally regulated G1. In certain postembryonic …


Control Of Developmental Timing In Caenorhabditis Elegans, Victor Ambros Jul 2000

Control Of Developmental Timing In Caenorhabditis Elegans, Victor Ambros

Victor R. Ambros

Studies of the nematode Caenorhabditis elegans have identified genetic and molecular mechanisms controlling temporal patterns of developmental events. Mutations in genes of the C. elegans heterochronic pathway cause altered temporal patterns of larval development, in which cells at certain larval stages execute cell division patterns or differentiation programs normally specific for other stages. The products of the heterochronic genes include transcriptional and translational regulators and two different cases of novel small translational regulatory RNAs. Other genes of the pathway encode evolutionarily conserved proteins, including a homolog of the Drosophila Period circadian timing regulator, and a member of the nuclear receptor …


The Lin-41 Rbcc Gene Acts In The C. Elegans Heterochronic Pathway Between The Let-7 Regulatory Rna And The Lin-29 Transcription Factor, Frank Slack, Michael Basson, Zhongchi Liu, Victor Ambros, H. Horvitz, Gary Ruvkun Mar 2000

The Lin-41 Rbcc Gene Acts In The C. Elegans Heterochronic Pathway Between The Let-7 Regulatory Rna And The Lin-29 Transcription Factor, Frank Slack, Michael Basson, Zhongchi Liu, Victor Ambros, H. Horvitz, Gary Ruvkun

Victor R. Ambros

Null mutations in the C. elegans heterochronic gene lin-41 cause precocious expression of adult fates at larval stages. Increased lin-41 activity causes the opposite phenotype, reiteration of larval fates. let-7 mutations cause similar reiterated heterochronic phenotypes that are suppressed by lin-41 mutations, showing that lin-41 is negatively regulated by let-7. lin-41 negatively regulates the timing of LIN-29 adult specification transcription factor expression. lin-41 encodes an RBCC protein, and two elements in the lin-413'UTR are complementary to the 21 nucleotide let-7 regulatory RNA. A lin-41::GFP fusion gene is downregulated in the tissues affected by lin-41 at the time that the let-7 …


Structure And Function Analysis Of Lin-14, A Temporal Regulator Of Postembryonic Developmental Events In Caenorhabditis Elegans, Yang Hong, Rosalind C. Lee, Victor R. Ambros Feb 2000

Structure And Function Analysis Of Lin-14, A Temporal Regulator Of Postembryonic Developmental Events In Caenorhabditis Elegans, Yang Hong, Rosalind C. Lee, Victor R. Ambros

Victor R. Ambros

During postembryonic development of Caenorhabditis elegans, the heterochronic gene lin-14 controls the timing of developmental events in diverse cell types. Three alternative lin-14 transcripts are predicted to encode isoforms of a novel nuclear protein that differ in their amino-terminal domains. In this paper, we report that the alternative amino-terminal domains of LIN-14 are dispensable and that a carboxy-terminal region within exons 9 to 13 is necessary and sufficient for in vivo LIN-14 function. A transgene capable of expressing only one of the three alternative lin-14 gene products rescues a lin-14 null mutation and is developmentally regulated by lin-4. This shows …


The Lin-4 Regulatory Rna Controls Developmental Timing In Caenorhabditis Elegans By Blocking Lin-14 Protein Synthesis After The Initiation Of Translation, Philip Olsen, Victor Ambros Dec 1999

The Lin-4 Regulatory Rna Controls Developmental Timing In Caenorhabditis Elegans By Blocking Lin-14 Protein Synthesis After The Initiation Of Translation, Philip Olsen, Victor Ambros

Victor R. Ambros

lin-4 encodes a small RNA that is complementary to sequences in the 3' untranslated region (UTR) of lin-14 mRNA and that acts to developmentally repress the accumulation of LIN-14 protein. This repression is essential for the proper timing of numerous events of Caenorhabditis elegans larval development. We have investigated the mechanism of lin-4 RNA action by examining the fate of lin-14 mRNA in vivo during the time that lin-4 RNA is expressed. Our results indicate that the rate of synthesis of lin-14 mRNA, its state of polyadenylation, its abundance in the cytoplasmic fraction, and its polysomal sedimentation profile do not …


The Timing Oflin-4rna Accumulation Controls The Timing Of Postembryonic Developmental Events Incaenorhabditis Elegans, Rhonda Feinbaum, Victor Ambros May 1999

The Timing Oflin-4rna Accumulation Controls The Timing Of Postembryonic Developmental Events Incaenorhabditis Elegans, Rhonda Feinbaum, Victor Ambros

Victor R. Ambros

The lin-4 gene encodes a small RNA that is required to translationally repress lin-14 toward the end of the first larval stage of Caenorhabditis elegans development. To determine if the timing of LIN-14 protein down-regulation depends on the temporal profile of lin-4 RNA level, we analyzed the stage-specificity of lin-4 RNA expression during wild-type development and examined the phenotypes of transgenic worms that overexpress lin-4 RNA during the first larval stage. We found that lin-4 RNA first becomes detectable at approximately 12 h of wild-type larval development and rapidly accumulates to nearly maximum levels by 16 h. This profile of …


Cell Cycle-Dependent Sequencing Of Cell Fate Decisions In Caenorhabditis Elegans Vulva Precursor Cells, Victor Ambros Apr 1999

Cell Cycle-Dependent Sequencing Of Cell Fate Decisions In Caenorhabditis Elegans Vulva Precursor Cells, Victor Ambros

Victor R. Ambros

In Caenorhabditis elegans, the fates of the six multipotent vulva precursor cells (VPCs) are specified by extracellular signals. One VPC expresses the primary (1 degrees ) fate in response to a Ras-mediated inductive signal from the gonad. The two VPCs flanking the 1 degrees cell each express secondary (2 degrees ) fates in response to lin-12-mediated lateral signaling. The remaining three VPCs each adopt the non-vulval tertiary (3 degrees ) fate. Here I describe experiments examining how the selection of these vulval fates is affected by cell cycle arrest and cell cycle-restricted lin-12 activity. The results suggest that lin-12 participates …


Developmental Regulation Of A Cyclin-Dependent Kinase Inhibitor Controls Postembryonic Cell Cycle Progression In Caenorhabditis Elegans, Yang Hong, Richard Roy, Victor Ambros Aug 1998

Developmental Regulation Of A Cyclin-Dependent Kinase Inhibitor Controls Postembryonic Cell Cycle Progression In Caenorhabditis Elegans, Yang Hong, Richard Roy, Victor Ambros

Victor R. Ambros

C. elegans cki-1 encodes a member of the CIP/KIP family of cyclin-dependent kinase inhibitors, and functions to link postembryonic developmental programs to cell cycle progression. The expression pattern of cki-1::GFP suggests that cki-1 is developmentally regulated in blast cells coincident with G1, and in differentiating cells. Ectopic expression of CKI-1 can prematurely arrest cells in G1, while reducing cki-1 activity by RNA-mediated interference (RNAi) causes extra larval cell divisions, suggesting a role for cki-1 in the developmental control of G1/S. cki-1 activity is required for the suspension of cell cycling that occurs in dauer larvae and starved L1 larvae in …


The Cold Shock Domain Protein Lin-28 Controls Developmental Timing In C. Elegans And Is Regulated By The Lin-4 Rna, Eric Moss, Rosalind Lee, Victor Ambros Mar 1997

The Cold Shock Domain Protein Lin-28 Controls Developmental Timing In C. Elegans And Is Regulated By The Lin-4 Rna, Eric Moss, Rosalind Lee, Victor Ambros

Victor R. Ambros

Mutations in the heterochronic gene lin-28 of C. elegans cause precocious development where diverse events specific to the second larval stage are skipped. lin-28 encodes a cytoplasmic protein with a cold shock domain and retroviral-type (CCHC) zinc finger motifs, consistent with a role for LIN-28 in posttranscriptional regulation. The 3'UTR of lin-28 contains a conserved element that is complementary to the 22 nt regulatory RNA product of lin-4 and that resembles seven such elements in the 3'UTR of the heterochronic gene lin-14. Both lin-4 activity and the lin-4-complementary element (LCE) are necessary for stage-specific regulation of lin-28. Deleting the LCE …


Heterochronic Genes, Victor Ambros Dec 1996

Heterochronic Genes, Victor Ambros

Victor R. Ambros

This chapter focuses on our current understanding of the C. elegans heterochronic gene pathway as determined by genetic characterization of the regulatory interactions among the genes (Ambros and Horvitz 1987; Ambros 1989; Liu and Ambros 1989) and recent progress on the cloning, sequencing, and molecular analysis of genes involved in this pathway (Ruvkun et al. 1989; Arasu et al. 1991; Papp et al. 1991; Wightman et al. 1991; Lee et al. 1993; Rougvie and Ambros 1995; E. Moss et al., in prep). Primary emphasis is placed on the developmental decisions controlled by the heterochronic genes, the regulatory interactions among genes …


Reversal Of Cell Fate Determination In Caenorhabditis Elegans Vulval Development, Susan Euling, Victor Ambros Jul 1996

Reversal Of Cell Fate Determination In Caenorhabditis Elegans Vulval Development, Susan Euling, Victor Ambros

Victor R. Ambros

In Caenorhabditis elegans, the fates of the multipotent vulval precursor cells (VPCs) are specified by intercellular signals. The VPCs divide in the third larval stage (L3) of the wild type, producing progeny of determined cell types. In lin-28 mutants, vulva development is similar to wild-type vulva development except that it occurs precociously, in the second larval stage (L2). Consequently, when lin-28 hermaphrodites temporarily arrest development at the end of L2 in the dauer larva stage, they have partially developed vulvae consisting of VPC progeny. During post-dauer development, these otherwise determined VPC progeny become reprogrammed back to the multipotent, signal-sensitive state …


Heterochronic Genes Control Cell Cycle Progress And Developmental Competence Of C. Elegans Vulva Precursor Cells, Susan Euling, Victor Ambros Mar 1996

Heterochronic Genes Control Cell Cycle Progress And Developmental Competence Of C. Elegans Vulva Precursor Cells, Susan Euling, Victor Ambros

Victor R. Ambros

Heterochronic genes control the timing of vulval development in the C. elegans hermaphrodite. lin-14 or lin-28 loss-of-function mutations cause the vulval precursor cells (VPCs) to enter S phase and to divide one larval stage earlier than in the wild type. A precocious vulva is formed by essentially normal cell lineage patterns, governed by the same intercellular signals as in the wild type. Mutations that prevent the normal developmental down-regulation of lin-14, activity delay or block VPC division and prevent vulval differentiation. A genetic pathway that includes lin-4, lin-14, and lin-28 controls when VPCs complete G1 and also controls when VPCs …


The Heterochronic Gene Lin-29 Encodes A Zinc Finger Protein That Controls A Terminal Differentiation Event In Caenorhabditis Elegans, Ann Rougvie, Victor Ambros Jul 1995

The Heterochronic Gene Lin-29 Encodes A Zinc Finger Protein That Controls A Terminal Differentiation Event In Caenorhabditis Elegans, Ann Rougvie, Victor Ambros

Victor R. Ambros

A hierarchy of heterochronic genes, lin-4, lin-14, lin-28 and lin-29, temporally restricts terminal differentiation of Caenorhabditis elegans hypodermal seam cells to the final molt. This terminal differentiation event involves cell cycle exit, cell fusion and the differential regulation of genes expressed in the larval versus adult hypodermis. lin-29 is the most downstream gene in the developmental timing pathway and thus it is the most direct known regulator of these diverse processes. We show that lin-29 encodes a protein with five zinc fingers of the (Cys)2-(His)2 class and thus likely controls these processes by regulating transcription in a stage-specific manner. Consistent …


The Caenorhabditis Elegans Heterochronic Gene Pathway Controls Stage-Specific Transcription Of Collagen Genes, Zhongchi Liu, Susan Kirch, Victor Ambros Jul 1995

The Caenorhabditis Elegans Heterochronic Gene Pathway Controls Stage-Specific Transcription Of Collagen Genes, Zhongchi Liu, Susan Kirch, Victor Ambros

Victor R. Ambros

In Caenorhabditis elegans, the terminal differentiation of the hypodermal cells occurs at the larval-to-adult molt, and is characterized in part by the formation of a morphologically distinct adult cuticle. The timing of this event is controlled by a pathway of heterochronic genes that includes the relatively direct regulatory gene, lin-29, and upstream genes lin-4, lin-14 and lin-28. Using northern analysis to detect endogenous collagen mRNA levels and collagen/lacZ reporter constructs to monitor collagen transcriptional activity, we show that the stage-specific switch from larval cuticle to adult cuticle correlates with the transcriptional activation of adult-specific collagen genes and repression of larval-specific …


Heterochronic Genes And The Temporal Control Of C. Elegans Development, Victor Ambros, Eric Moss Mar 1994

Heterochronic Genes And The Temporal Control Of C. Elegans Development, Victor Ambros, Eric Moss

Victor R. Ambros

The heterochronic genes of Caenorhabditis elegans encode part of a regulatory system that controls the temporal component of cell fates in development. The genes have been characterized genetically and molecularly, and their study has so far revealed a genetic hierarchy that specifies sequences of developmental events, a novel RNA-mediated mechanism of gene regulation and a reprogramming phenomenon associated with arrested development.


The C. Elegans Heterochronic Gene Lin-4 Encodes Small Rnas With Antisense Complementarity To Lin-14, Rosalind Lee, Rhonda Feinbaum, Victor Ambros Dec 1993

The C. Elegans Heterochronic Gene Lin-4 Encodes Small Rnas With Antisense Complementarity To Lin-14, Rosalind Lee, Rhonda Feinbaum, Victor Ambros

Victor R. Ambros

lin-4 is essential for the normal temporal control of diverse postembryonic developmental events in C. elegans. lin-4 acts by negatively regulating the level of LIN-14 protein, creating a temporal decrease in LIN-14 protein starting in the first larval stage (L1). We have cloned the C. elegans lin-4 locus by chromosomal walking and transformation rescue. We used the C. elegans clone to isolate the gene from three other Caenorhabditis species; all four Caenorhabditis clones functionally rescue the lin-4 null allele of C. elegans. Comparison of the lin-4 genomic sequence from these four species and site-directed mutagenesis of potential open reading frames …


Efficient Gene Transfer In C.Elegans: Extrachromosomal Maintenance And Integration Of Transforming Sequences, Craig Mello, James Cramer, Dan Stinchcomb, Victor Ambros Nov 1991

Efficient Gene Transfer In C.Elegans: Extrachromosomal Maintenance And Integration Of Transforming Sequences, Craig Mello, James Cramer, Dan Stinchcomb, Victor Ambros

Victor R. Ambros

We describe a dominant behavioral marker, rol-6(su-1006), and an efficient microinjection procedure which facilitate the recovery of Caenorhabditis elegans transformants. We use these tools to study the mechanism of C.elegans DNA transformation. By injecting mixtures of genetically marked DNA molecules, we show that large extrachromosomal arrays assemble directly from the injected molecules and that homologous recombination drives array assembly. Appropriately placed double-strand breaks stimulated homologous recombination during array formation. Our data indicate that the size of the assembled transgenic structures determines whether or not they will be maintained extrachromosomally or lost. We show that low copy number extrachromosomal transformation can …


Alternative Temporal Control Systems For Hypodermal Cell Differentiation In Caenorhabditis Elegans, Zhongchi Liu, Victor Ambros Mar 1991

Alternative Temporal Control Systems For Hypodermal Cell Differentiation In Caenorhabditis Elegans, Zhongchi Liu, Victor Ambros

Victor R. Ambros

Beginning of article: In certain multicellular organisms, genetic regulatory systems that specify the timing of cell division, differentiation and morpho-genesis must accommodate environmental and physiological contingencies that perturb or arrest development. For example, Caenorhabditis elegans can either develop continuously through four larval stages (L1–L4) or arrest indefinitely as a 'dauer larva' at the second larval (L2) moult, and later resume L3 and L4 development. At the larva-to-adult (L4) moult of both con-tinuous and 'post-dauer' development, hypodermal cells switch (the 'L/A switch') from a proliferating state to the terminally differentiated state. Four temporal regulators, lin-4, lin-14, lin-28 and lin-29, have been …