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Improved CRISPR/Cas9 knock-in efficiency via the self-excising cassette (SEC) selection method in C. elegans.

George Huang, Bailey de Jesus, Alex Koh, Sara Blanco, Aubrie Rettmann, Ella DeMott, Melynda Sylvester, Cassie Ren, Carrie Meng, Skye Waterland, Anita Rhodes, Persephone Alicea, Abbey Flynn, Daniel J Dickinson, Ryan Doonan,

Streamlined, selection-based CRISPR knock-in protocols for <i>C. elegans</i> were first introduced six years ago (Dickinson <i>et al</i>. 2015; Schwartz and Jorgensen 2016). Though these selection-based approaches are powerful, one drawback has been the requirement to inject large numbers of P0 worms (~30-60 per gene target). We have found that a ... Read more >>

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[2021, 2021:]

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Missense mutation of a conserved residue in UNC-112 (kindlin) eliminates binding to PAT-4 (ILK).

Hiroshi Qadota, Andres F Oberhauser, Guy M Benian,

<i>C. elegans</i> UNC-112 (kindlin) is required for muscle sarcomere assembly, and is one component of a conserved four-protein complex that associates with the cytoplasmic tail of integrin at the base of integrin adhesion complexes in muscle. The four-protein complex consists of UNC-112 (kindlin), PAT-4 (integrin linked kinase; ILK), PAT-6 (alpha-parvin), ... Read more >>

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[2021, 2021:]

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Deletion of a putative HDA-1 binding site in the hlh-2 promoter eliminates expression in C. elegans dorsal uterine cells.

Taylor N Medwig-Kinney, Nicholas J Palmisano, David Q Matus,

The helix-loop-helix transcription factor <i>hlh-2</i> (E/Daughterless) has been shown to play an important role in regulating cell fate patterning, cell cycle, and basement membrane invasion in the context of the development of the <i>C. elegans</i> somatic gonad. Here, using CRISPR/Cas9 genome engineering, we generated a new <i>hlh-2</i> allele (<i>hlh-2(Δ-1303-702)</i>) in ... Read more >>

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[2021, 2021:]

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Modeling human glucose-6-phosphate dehydrogenase mutations using C. elegans GSPD-1.

Luiza N Loges, Katherine M Walstrom,

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked, recessive condition that causes intermittent jaundice or hemolytic anemia because of low NADPH levels in red blood cells. We performed steady-state enzyme kinetics with the recombinant <i>C. elegans</i> ortholog of human G6PD, GSPD-1, and two mutants containing amino acid changes found in human ... Read more >>

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[2021, 2021:]

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Paternal mitochondria from an rmd-2, rmd-3, rmd-6 triple mutant are properly positioned in the C. elegans zygote.

Iris Y Juanico, Christina M Meyer, John E McCarthy, Ting Gong, Francis J McNally,

RMD-1,2,3,6 (regulator of microtubule dynamics) is a family of homologous proteins conserved between humans and <i>C. elegans</i>. Human RMD-3/PTPIP51 is a mitochondrial protein that tethers mitochondria to the endoplasmic reticulum. <i>C. elegans</i> RMD-2, 3, and 6 are expressed in sperm. To test whether paternal RMD-2, 3, 6 might redundantly tether ... Read more >>

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[2021, 2021:]

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The MLK-1/SCD-4 Mixed Lineage Kinase/MAP3K functions to promote dauer formation upstream of DAF-2/InsR.

Neal R Rasmussen, Harold E Smith, David J Reiner,

The <i>C. elegans</i> dauer is an alternative third stage larva induced by dense population and adverse environmental conditions. Genes whose mutants caused dauer formation constitutive (Daf-c) and dauer formation defective (Daf-d) phenotypes were ordered via epistasis into a signaling network, with upstream DAF-7/TGF-beta and DAF-11/receptor guanylyl cyclase defining sensory branches ... Read more >>

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[2021, 2021:]

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Genetic Mapping of a new Hippo allele, HpoN.1.2, in Drosophila melanogaster.

Jamie L Siders, Kayla L Bieser, Danielle R Hamill, Erika C Acosta, Olivia K Alexander, Humza I Ali, Micah J Anderson, Hayden R Arrasmith, Mustafa Azam, Nikki J Beeman, Hassan Beydoun, Lauren J Bishop, Morgan D Blair, Brianna Bletch, Heather R Bline, Jennifer C Brown, Kelly M Burns, Karina C Calagua, Lexie Chafin, William Ah Christy, Carlyn Ciamacco, Hannah Cizauskas, Caitlyn M Colwell, Abigail R Courtright, Lucero Diaz Alavez, Rayne Is Ecret, Fatima Edriss, Taylor G Ellerbrock, Madison M Ellis, Erica M Extine, Eric Feldman, Luke J Fickenworth, Caroline M Goeller, Alexis S Grogg, Yailine Hernandez, Abigail Hershner, Megan M Jauss, Leyre Jimenez Garcia, Katey E Franks, Ethan T Kazubski, Emily R Landis, Jon Langub, Tia N Lassek, Triet C Le, Julia M Lee, Daniel P Levine, Phoebe J Lightfoot, Natasha Love, Ali Maalhagh-Fard, Colin Maguire, Brynna E McGinnis, Bhargavi V Mehta, Veronica Melendrez, Zimri E Mena, Seth Mendell, Petra Montiel-Garcia, Autumn S Murry, Riley A Newland, Ryan M Nobles, Neha Patel, Yashodhara Patil, Cassidy L Pfister, Victoria Ramage, Mya R Ray, Joseph Rodrigues, Victoria C Rodriquez, Yara Romero, Alexandra M Scott, Nicholas Shaba, Samantha Sieg, Kayla Silva, Sahiba Singh, Aleksandria J Spargo, Savanna J Spitnale, Nicole Sweeden, Logan Tague, Breanna M Tavernini, Kathleen Tran, Liselle Tungol, Kylie A Vestal, Amber Wetherbee, Kayla M Wright, Anthony T Yeager, Rehab Zahid, Jacob D Kagey,

Genetic screens provide a mechanism to identify genes involved with different cellular and organismal processes. Using a Flp/FRT screen in the <i>Drosophila</i> eye we identified mutations that result in alterations and de-regulation of cell growth and division. From this screen a group of undergraduate researchers part of the Fly-CURE consortium ... Read more >>

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[2021, 2021:]

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Deletion of the RGD motif in LON-2/glypican is associated with morphological abnormalities.

Aileen Park, Zhongqiang Qiu, Myeongwoo Lee,

The <i>lon-2</i> gene in <i>Caenorhabditis elegans</i> encodes a heparan sulfate proteoglycan family glypican that negatively regulates the BMP signaling pathway responsible for controlling body length. LON-2 contains multiple functional domains, including an RGD (Arg-Gly-Asp) motif at amino acid number from 348 to 350. A novel mutant allele of <i>lon-2</i> was ... Read more >>

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[2021, 2021:]

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CRISPR-edit point mutant allele detection (CEPAD)-PCR method for rapid screening of CRISPR edited point mutations.

Kenneth Trimmer, Swathi Arur,

CRISPR-Cas9 mediated genome editing is widely used for generating genetic lesions in <i>C. elegans</i>. Detection of single-site mutations in F1 progeny after CRISPR-Cas9 injections is currently labor intensive due to lack of a single step PCR-based detection method. Here we present CEPAD-PCR, an allele-specific PCR detection method based on generating ... Read more >>

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[2021, 2021:]

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Serum-free adapted Drosophila S2R+ line is amenable to RNA interference.

Arthur Luhur, Daniel Mariyappa, Kristin M Klueg, Stephen L Rogers, Andrew C Zelhof,

We have previously adapted a select number of <i>Drosophila</i> cell lines to grow in serum-free media supplemented with fly extract. This condition is arguably more representative of a native growth environment. Here, we validated that the fly extract adapted line, S2R+ (FEx 2.5%) is amenable to RNAi. RNAi against <i>Rho1</i> ... Read more >>

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[2021, 2021:]

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CRISPRcruncher: A tool for engineering restriction sites into coding regions.

Samuel F Fay, David S Fay, Vikram E Chhatre,

CRISPR/Cas9 genome editing strategies often rely on the placement of an introduced restriction endonuclease (RE) site adjacent to the genomic edit of interest. This allows for rapid initial PCR-based detection of cells and organisms containing the edit of interest and may also be used for subsequent genotyping. Nevertheless, engineering RE ... Read more >>

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[2021, 2021:]

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C. elegans spermatozoa lacking spe-45 are incapable of fusing with the oocyte plasma membrane.

Jun Takayama, Tatsuya Tajima, Shuichi Onami, Hitoshi Nishimura,

<i>C. elegans spe-9</i> class genes encode sperm proteins with indispensable roles during fertilization. We have previously reported that <i>spe-45</i> belongs to the <i>spe-9</i> class, based on the finding that self-sperm of <i>spe-45(tm3715)</i> hermaphrodites were not consumed by fertilization. In this study, we directly observed live fertilization in the spermatheca of ... Read more >>

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[2021, 2021:]

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Conserved missense variant in ALDH1A3 ortholog impairs fecundity in C. elegans.

Wan-Rong Wong, Shayda Maher, Jun Young Oh, Katherine I Brugman, Shahla Gharib, Paul W Sternberg,

Accumulating evidence demonstrates that mutations in <i>ALDH1A3</i> (the aldehyde dehydrogenase 1 family, member A3) are associated with developmental defects. The ALDH1A3 enzyme catalyzes retinoic acid biosynthesis and is essential to patterning and neuronal differentiation in the development of embryonic nervous system. Several missense mutations in <i>ALDH1A3</i> have been identified in ... Read more >>

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[2021, 2021:]

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A histamine-gated channel is an efficient negative selection marker for C. elegans transgenesis.

Sonia El Mouridi, Sarah AlHarbi, Christian Frøkjær-Jensen,

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[2021, 2021:]

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Comparison of N- and C-terminally endogenously GFP-tagged WEE-1.3 strains in C. elegans.

Lourds M Fernando, Kyrionna Golliday, Ruby Boateng, Anna K Allen,

We have generated a WEE-1.3 strain in <i>C. elegans</i> wherewe have endogenously tagged the C-terminus with GFP<i>.</i> In this publication we demonstrate that this new strain exhibits the same expression localization pattern as the WEE-1.3 antibody and N-terminally endogenously GFP-tagged WEE-1.3 strain that have been previously published. We also show ... Read more >>

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[2021, 2021:]

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Genetic analysis of the interaction between the N- and C-terminal halves of UNC-112 (kindlin).

Hiroshi Qadota, Yating Luo, Andres F Oberhauser, Guy M Benian,

MicroPubl Biol (microPublication biology)
[2020, 2020:]

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Expression of an endosome-excluded Cd63 prevents axis elongation in Xenopus.

Jennifer Kreis, Ramona Bonß, Kerstin Feistel, Philipp Vick,

MicroPubl Biol (microPublication biology)
[2020, 2020:]

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Engineering essential genes with a "jump board" strategy using CRISPR/Cas9.

Ye Duan, Sungwook Choi, Charles Nelson, Victor Ambros,

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[2020, 2020:]

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In vitro screening methods for parasites: the wMicroTracker & the WormAssay.

Emma Gunderson, Christina Bulman, Mona Luo, Judy Sakanari,

MicroPubl Biol (microPublication biology)
[2020, 2020:]

Cited: 2 times

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Plant hexokinase phylogenetic analysis highlights a possible regulation by the posttranslational modifier SUMO.

Pedro Humberto Castro, Sara Freitas, Herlander Azevedo,

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[2020, 2020:]

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Arabidopsis thaliana growth is independently controlled by the SUMO E3 ligase SIZ1 and Hexokinase 1.

Pedro Humberto Castro, Nuno Verde, Herlander Azevedo,

MicroPubl Biol (microPublication biology)
[2020, 2020:]

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Codon optimized Tol2 transposase results in increased transient expression of a crystallin-GFP transgene in zebrafish.

Allison S Mackey, Priscilla S Redd, April DeLaurier, C Nathan Hancock,

MicroPubl Biol (microPublication biology)
[2020, 2020:]

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Novel actions of arecoline in the C. elegans motor circuit.

Katherine A McCulloch, Yishi Jin,

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[2020, 2020:]

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A partial T-DNA insertion near KNAT1 results in lobed Arabidopsis thaliana leaves.

Karah Moulton, Stephanie Diaz, Ashley Strother, C Nathan Hancock,

MicroPubl Biol (microPublication biology)
[2020, 2020:]

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Two alleles of unc-52 locus disrupting potential cell-binding motif of UNC-52.

Rachel Wilsey, Sabrina Hodge, Krysta Kenney, Jacob Wahl, Roshni Jaffery, Avery Brau, Zhongqiang Qiu, Myeongwoo Lee,

MicroPubl Biol (microPublication biology)
[2020, 2020:]

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