UNIVERSITY OF MASSACHUSETTS
This ARRA award is an administrative supplement to NIH Grant R01 GM081422. The stated goals of the supplement request were two-fold. First, we proposed to develop a new high-throughput sequencing / in vitro selection method in order to increase our capacity to measure RNA-binding protein specificity, focusing on proteins that regulate nematode maternal gene expression. Second, we proposed to make use of a powerful new single copy transgene integration method to make fluorescent reporter strains that enhance our ability to analyze the function of putative cis-regulatory sequences in live worms. The expected outcome is an increase the pace of research proposed in NIH Grant R01 GM081422, and the retention of a talented post-doctoral research associate to perform the described experiments.
| AWARD OVERVIEW |
| Award Number |
3R01GM081422-02S1 |
Funding Agency |
Department of Health and Human Services |
| Total Award Amount |
$124,975 |
Project Location - City |
Worcester |
| Award Date |
01/08/2010 |
Project Location - State |
MA |
| Project Status |
Completed |
Project Location - Zip |
01655-0002
|
| Jobs Reported |
0.14 |
Congressional District |
03 |
| Project Location - Country |
US |
|
|
Recipient Information
(Grants)
| Recipient Information (Grants) |
|
Recipient Name
|
UNIVERSITY OF MASSACHUSETTS |
| Recipient DUNS Number |
603847393
|
| Recipient Address |
55 LAKE AVE N |
| Recipient City |
WORCESTER |
| Recipient State |
Massachusetts |
| Recipient Zip |
01655-0002 |
| Recipient Congressional District |
03 |
| Recipient Country |
USA |
Required to Report Top 5
Highly Compensated Officials |
No |
Projects and Jobs Information
| Projects and Jobs Information |
| Project Title |
ARRA-RNA recognition by maternal gene silencers in nematodes |
| Project Status |
Completed |
| Final Project Report Submitted |
Yes |
| Project Activities Description |
Highway, Street, and Bridge Construction |
| Quarterly Activities/Project Description |
In this quarter, we refined our method to rapidly generate C. Elegans lines that express 3-UTR reporter transgenes. In the previous quarter, we showed that the single-copy integration (MosSCI) approach of Jorgensen and coworkers could be adapted to generate multiple reporter lines simultaneously. In brief, we discovered that a library of reporter constructs could be injected into a single animal and that individual reporter lines can be generated by stochastic integration of members of the library during the integration step. Our approach minimizes the effort required to microinject and identify rescued lines with individual reporters. Moreover, a 'producer' line is generated that can be stored indefinitely, revived when needed, and induced over and over again to generate new single copy integrants. This quarter, we have devised a simple PCR based assay to facilitate identification of integrants from the progeny from the producer strain. We have now cloned 45 3-UTR reporter constructs, and combined 30 of these into one library to see what fraction of the reporters can be recovered from a single animal. We have hit a minor stumbling block caused by epigenetic germline-specific transgene silencing. We are currently trying to back-cross our integrated lines to alleviate silencing. We also plan to passage our lines on RNAi food that inhibits transgene silencing. In the near future, we plan to increase the scope of the project to include promoter reporters and 3-UTR reporters that are expressed in the soma. We have also made progress in determining RNA binding protein specificity using high-throughput sequencing. We have completed multiple rounds of SELEX with OMA-1, a CCCH-type zinc finger protein. We will soon prepare libraries from each round of SELEX in a format suitable for Illumina sequencing. We hope to map the trajectory of the in vitro selection across multiple rounds in order to identify the minimal number of rounds. |
| Jobs Created |
0.14 |
| Description of Jobs Created |
Post Doc Assoc |
Purchaser Information
(Grants)
| Purchaser Information |
| Contracting Office ID |
Not Reported |
| Contracting Office Name |
Not Available |
| Contracting Office Region |
Not Available |
| TAS Major Program |
75-0852 |
| Award Information |
| Award Date |
01/08/2010 |
| Award Number |
3R01GM081422-02S1 |
| Order Number |
|
| Award Type |
Grants |
| Funding Agency ID |
75 |
| Funding Agency Name |
Department of Health and Human Services |
| Funding Office Name |
Not Available |
| Awarding Agency ID |
75 |
| Awarding Agency Name |
Department of Health and Human Services |
| Amount of Award |
$124,975 |
| Funds Invoiced/Received |
$124,975 |
| Expenditure Amount |
$124,975 |
| Infrastructure Expenditure Amount |
$0 |
| Infrastructure Purpose and Rationale |
Not Reported |
| Infrastructure Point of Contact Name |
Not Reported |
| Infrastructure Point of Contact Email |
Not Reported |
| Infrastructure Point of Contact Phone |
Not Reported |
| Infrastructure Point of Contact Address |
Not Reported |
| Infrastructure Point of Contact City |
Not Reported |
| Infrastructure Point of Contact State |
Not Reported |
| Infrastructure Point of Contact Zip |
Not Reported |
Product or Service Information
(Grants)
| Product or Service Information |
| Primary Activity Code |
237310 |
| Activity Description |
Highway, Street, and Bridge Construction |
| Sub-Awards Information |
| Sub-awards to Organizations |
0 |
| Sub-award Amounts to Organizations |
$0 |
| Sub-Awards to Individuals |
0 |
| Sub-Award Amounts to Individuals |
$0 |
| Number of Sub-awards less than $25,000/award |
0 |
| Amount of Sub-awards less than $25,000/award |
$0 |
| Number of payments to vendors greater than $25,000 |
0 |
| Total Amount of payments to vendors greater than $25,000/award |
$0 |
| Number of payments to vendors less than $25,000/award |
11 |
| Total Amount of payments to vendors less than $25,000/award |
$12,184 |
| Location Information |
| Latitude, Longitude |
42º 16' 39",
-71º 45' 33" |
| Congressional District |
03 |
| Address 1 |
55 Lake Avenue North |
| Address 2 |
|
| City |
Worcester |
| County |
Worcester |
| State |
MA |
| Zip |
01655-0002 |
|
 |