- LMC Special Volunteer or Guest Researcher Appt. Checklist (New/Renewal) - Non-U.S. Citizen Word(33KB) , PDF(11KB)
- LMC Post-Doc IRTA, Appt. Renewal Checklist Word(32KB) , PDF(10KB)
- LMC Post-Doc IRTA, New Appt. Checklist Word(32KB) , PDF(10KB)
- LMC Post-Doc Visiting Fellow, Appt. Renewal Checklist, Word(49KB) , PDF(66KB)
- LMC Post-Doc Visiting Fellow, New Appt. Checklist Word(44KB) , PDF(60KB)
- LMC Research Fellow (VP), New Appt. Checklist Word(51KB) , PDF(84KB)
- LMC Research Fellow (VP), Appt. Renewal Checklist Word(49KB) , PDF(66KB)
- LMC Special Volunteer or Guest Researcher Appt. Checklist (New/Renewal) - U.S. Citizen Word(33KB) , PDF(10KB)
Lackford and Scruggs Recognized for Best Posters at NIEHS Science Day
We want to congratulate LMC members Brad Lackford and Ben Scruggs for being awarded best posters at the recent NIEHS Science Day. Brad is a biologist with Dr. Guang Hu, and his poster described their study of Fip1, an mRNA 3’ processing factor belonging to the CPSF complex that is indispensible for the maintenance of mouse embryonic stem cell (mESC) self-renewal and pluripotency as well as somatic cell reprogramming. Fip1 exerts it control, in part, by maintaining an ES cell specific alternate polyadelnylation (APA) program. Mechanistically, Fip1 acts through the recognition of a suboptimal poly-adenylation site to produce mRNAs with a shorter 3’ UTR. Their data show a specific biological significance to APA as a novel model for the post-transcriptional regulation of self-renewal and pluripotency in mESCs.
Ben is a post doctoral fellow in Dr. Karen Adelman’s lab. Ben’s poster described his studies of the pausing of RNA Pol II during early transcriptional elongation, a known critical regulatory mechanism for RNA synthesis. Consistent with growing evidence that many enhancers are sites of Pol II transcription, they find a considerable portion of 5’-capped RNAs (scRNA) aligning to experimentally validated enhancer regions. They then utilized scRNAs to define extragenic transcription start sites (eTSSs) in an unbiased fashion. These eTSSs overlap considerably with experimentally validated enhancers. They also found the transcriptional activator GAGA at eTSSs with enhancer features, along with its known binding partner, the ISWI chromatin remodeler. These data provide new insights into transcriptional regulation at enhancers and highlight the utility of scRNA sequencing to define TSSs.
Fellows Award for Research Excellence for 2013
Congratulations to LMC members (l-r) Dr. George Fromm and Dr. Senthil Cinghu on each receiving a Fellows Award for Research Excellence for 2013. Their abstracts were selected from numerous entries from post doctoral fellows throughout the NIH. Dr. Fromm works in the laboratory of Dr. Karen Adelman and his abstract described their studies on a critical point of gene regulation that occurs during early transcription elongation, when pause-inducing factors such as the NELF complex cause RNA polymerase II to halt just downstream of the promoter. Dr. Fromm has used a knockout mouse model to demonstrate that NELF-mediated pausing plays a critical role in establishing MAPK/ERK signaling pathway activity during development. Dr. Cinghu works in the laboratory of Dr. Raja Jothi. His abstract described their efforts to understand the precise molecular mechanisms underlying the balance between self-renewal and differentiation. Using a systematic meta-analysis of published gene expression data, they identified a novel mechanism involving a Nucleolin-dependent bistable switch regulating the homeostatic balance between self-renewal and differentiation in embryonic stem cells.
Cold Spring Harbor Asia Fellowship awarded to Dr. Wang
Dr. Li Wang, a Visiting Fellow in the Stem Cell Biology Group led by Dr. Guang Hu, was recently awarded a Cold Spring Harbor Asia Fellowship in recognition of his outstanding poster presentation at the Cold Spring Harbor Asia meeting on Stem Cells and Developmental Mechanisms. The meeting was held in early December, 2012 in Suzhou, China. Dr. Wang's research is focused on how to improve cardiac differentiation efficiency through genetical modification of specific genes in human embryonic stem cells (ESCs). He found that Cnot3, a pluripotent factor identified by the Hu lab, plays a critical role during mouse heart development and promotes human cardiac differentiation in vitro. Thus, Cnot3 may serve as a potential target to overcome the low efficiency of cardiac cell differentiation and improve the outcome of stem cell replacement therapy in heart disease.
Available Techniques & Expertise in LMC
Sequencing Lab Group (SLG) Mission
The mission of the Automated DNA Sequencing Core Unit is to provide excellent quality data in a reasonable amount of time. Our policy is to provide each investigator an equal opportunity to submit samples and our goal is to return analyzed data within 1 to 2 days from the samples submission. Our average turnaround time is less than 24 hours from sample submission. The NIEHS DNA Sequencing Core uses biochemical methods to determine the order of nucleotide bases - adenine, guanine, cytosine and thymine - in a DNA sample. The core uses Perkin-Elmer ABI Model 3100 and Model 3130 sequencers, which offer a variety of services including single pass primer extension using plasmid DNA, PCR products or genomic DNA as a sequencing template. The single pass extension using conventional sequencing primers such as M13, T7, T3 and SP6 or a custom primer, can provide 600-650 base pairs of "good" or viable sequence data with a quick turnaround time. Shotgun sequencing strategies can be used to sequence a variety of samples including cosmids, BACs and PACs.
The DNA Sequencing Core does not accept samples from the general public, but any investigator from within the Institute may submit samples for analysis at no cost. Greg Solomon manages the facility, with biologist Jason Malphurs and sequencing engineer John Otstot assisting in the generation of sequence data.
Sequencing Lab Group (SLG) Publications(20KB)
Sequencing Lab Group (SLG) Staff
Tel (919) 541-1812
Tel (919) 316-4588
Tel (919) 541-0493
Nicole Reeves, M.S.
Tel (919) 541-0476
In Vivo Imaging System for Bioluminescence and Fluorescence
Bioluminescent Images of Tumor-bearing Mice
Human breast carcinoma cells expressing firefly luciferase were injected into the mammary fat pads of nude (immunocompromised) female mice. To visualize the location of tumor cells, mice were injected intraperitoneally with luciferin, then anesthetized and placed in the Caliper Spectrum in vivo imaging system at NIEHS. Each image shows an overlay of the bioluminescent signal on a white light image of the mouse (ventral view).
A) Mouse showing bioluminescent signal from the primary tumor in the mammary fat pad 6 weeks after injection of tumor cells. Note the scale indicates the most intense pixels represent approximately 1 x 109 photons/sec/cm2.
B) Same mouse, imaged one week after surgical resection of the primary tumor, showing three areas of bioluminescence: 1) potential remaining cells from primary tumor, or metastasis to adjacent fat pad; 2) potential metastasis to another mammary fat pad or axillary lymph nodes; and 3) potential lung metastasis. Maximal bioluminescent signal in this mouse was approximately 1 x 104 photons/sec/cm2 which would not have been visible when the primary tumor was present.
C) Another mouse imaged one week after the primary tumor was surgically resected. Note the absence of tumor over the mammary fat pad, but the appearance of bioluminescence over the chest. The most intense signal from this apparent lung metastasis represents approximately 4 x 104 photons/sec/cm2. (D. Ray, P. Myers, and J. Roberts)
Contact John Roberts in LMC for information and training in the use of the Caliper Spectrum IVIS located within the animal facility at NIEHS.
Calendar of Events
For more on the LMC Journal Club and Seminar Series, see below.
Journal Club Mission
The LMC journal club has been established to foster scientific discussion on the most recent science publications of interest to the laboratory. All members are invited to pick, present and lead a discussion on a recent journal article.
Notes to Participants
The 2013-2014 journal club will be held on the 2rd Wednesday of each month at noon unless otherwise notified. The meeting dates and locations are listed below. Anyone in the LMC is welcome to present, including fellows, biologists and principle investigators. Anyone interested in leading a discussion should contact Hrisavgi (Chrys) Kondilis-Mangum at (919) 541-5186 or by email at email@example.com.
We ask that a PDF file and a short description of the article to be discussed to be emailed to the organizer for distribution at least 7 days prior to leading the meeting. You will be sent a reminder two weeks before. We also ask if a speaker must cancel their discussion that reasonable notice is given so that the organizer can locate a substitute or rearrange the schedule.
2013-2014 Journal Club Schedule
|2013-2014 Meeting Dates||Room||Presenter|
|November 13||D450||Eric Milliman|
|December 11||D450||Senthikumar Cinghu|
|January 08||D450||Ben Scruggs|
|February 12||D450||Motoki Takaku|
|March 12||D450||Andrew Oldfield|
|April 09||D450||Nolan Gokey|
|May 14||D450||Beth Bowman|