G
Cell Line: HEK293
Cell Line Supplier: Synthego
Species: Human
Gene Name: POLR3B
Modification Type: SNP change
Modification details: 1625 A>G
Type of edit: Homozygous
Additional deliverables/requests:
Requesting 2 homozygous clones + 2 additional clones
that are heterozygous
2.Creation of knockin pool (small) - Qty (1) EA
3.Additional clones - heterozygous. - Qty (2) EA
4.POLR3E T275M Clones in HEK293 - Qty (1) EA
Cell Line: HEK293
Cell Line Supplier: Synthego
Species: Human
Gene Name: POLR3E
Modification Type: SNP change
Modification details:
824C>T (T275M)
Type of edit: Homozygous
Additional deliverables/requests:
Requesting 2 homozygous clones + 2 additional clones
that are heterozygous
5. Creation of knockin pool (small) - Qty (1) EA
6.Additional Clones - Heterozygous - Qty (2) EA
Project Milestones
Milestone 1a - Design and Synthesis of modified sgRNA and donor template
Communication: Completion of design & beginning of synthesis
Milestone 1b - Transfection and genotyping of pool
Communication: Editing efficiency of the pool
Milestone 2 - Clonal isolation and initial genotyping
Communication: Genotype of the identified clones
Milestone 3 - Sequence validation of alleles
Communication: Final Report
Project Deliverables
2 vials each of two homozygous POLR3B clones of edited cells at 1 x 10^6 cells/vial containing the mutation N542S
2 vials each of two heterozygous POLR3B clones of edited cells at 1 x 10^6 cells/vial containing the mutation N542S
4 vials of Wild-Type Cells at 1 x 10^6 cells/vial.
2 vials each of two homozygous POLR3E clones of edited cells at 1 x 10^6 cells/vial containing the mutation T275M
2 vials each of two heterozygous POLR3E clones of edited cells at 1 x 10^6 cells/vial containing the mutation t275m
Final project summary document.
Certificate of Analysis for the delivered cells, including mycoplasma data.
The service will create two different immortal HEK293 cell lines that will be models for two patient mutations. They each will have different mutations in RNA Pol III, the RNA polymerase that synthesizes all transfer RNAs (tRNAs). One cell line will have a mutation in POLR3E subunit, the other one in the POLR3B subunit. The patient that has a mutation POLR3B has a mutation in a different, unrelated gene and we want to demonstrate the phenotype we see in the patient's cells is really due to the POLR3B mutation.
Synthego has extensive experience in creating specific point mutations in HEK293 cells and can deliver the requested edited cells. Synthego is the only provider to exclusively utilize sgRNA and Cas9 protein to form an RNP to perform editing, which leads to higher rates of success, shorter timeframes and lower cost. Synthego provides a 100% money back guarantee.
Synthego synthetic sgRNA is designed and utilized in a ribonucleoprotein (RNP) to alter cells. Synthego's synthetic sgRNA is manufactured via a proprietary methodology created and utilized by Synthego. Synthego utilizes a proprietary screening strategy that images the growing clones every two days to ensure single cell clonal growth is achieved and only single cell clones are selected for further expansion and genotyping.
The proposed contract action is for supplies or services for which the Government intends to solicit and negotiate with only one source under authority of the 41 U.S.C. 253(c) (1), FAR 6.302. Only one responsible source and no other supplies or services will satisfy agency requirements. The offeror must include a completed copy of the provision of FAR Clause 52.212-3, Offeror Representations and Certifications - Commercial Items with its offer. The provisions of FAR Clause 52.212-1 Instructions to Offerors - Commercial Items; FAR Clause 52.212-2, Evaluation - Commercial Items; FAR Clause 52.212-4, Contract Terms and Conditions - Commercial Items; and FAR 52.212-5, Contract Terms and Conditions Required to Implement Statutes or Executive Orders - Commercial Items - Deviation for Simplified Acquisitions applies to this acquisition. The offeror must include their Dun & Bradstreet Number (DUNS), the Taxpayer Identification Number (TIN), and the certification of business size. The clauses are available in full text at http://www.acquisition.gov/far/. Responses to this solicitation must include sufficient information to establish the interested parties' bona-fide capabilities of providing all services and or products specified in this synopsis should submit a copy of their quotation to the below address or via email to [email protected]. Offers must also be accompanied by descriptive literature, delivery timeframe, warranties and/or other information that demonstrates that the offer meets all the foregoing requirements. Quotations will be due fifteen calendar days from the publication date of this synopsis or by August 15, 2019 by 9 am, EST. via email or postal mail. The quotation must reference "Solicitation number" NICHD-19-181. Quotations sent by postal mail or other mailing services must be submitted to the following address: Eunice Kennedy Shriver National Institute of Child Health and Human Development, 6710B Rockledge Drive, MSC 1159B, Bethesda, MD 20892. Attention: Tina Robinson, by the date and time mentioned above. Any questions must be sent via email to [email protected] and must include solicitation# NICHD-19-181 in the subject line of email. Faxed copies/responses will not be accepted.
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