Tuneable endogenous mammalian target complementation via multiplexed plasmidbased recombineering

Violeta Beltran-Sastre; Hannah Benistry; Julia Burnier; Imre Berger; Luis Serrano; Christina Kiel

doi:10.1038/srep17432

Tuneable endogenous mammalian target complementation via multiplexed plasmidbased recombineering

Violeta Beltran-Sastre, Hannah Benistry, Julia Burnier, Imre Berger, Luis Serrano, Christina Kiel

School of Biochemistry

Research output: Contribution to journal › Article (Academic Journal) › peer-review

2 Citations (Scopus)

287 Downloads (Pure)

Abstract

Understanding the quantitative functional consequences of human disease mutations requires silencing of endogenous genes and expression of mutants at close to physiological levels. Changing protein levels above or below these levels is also important for system perturbation and modelling. Fast design optimization demands flexible interchangeable cassettes for endogenous gene silencing and tuneable expression. Here, we introduce ‘TEMTAC’, a multigene recombineering and delivery system for simultaneous siRNA-based knockdown and regulated mutant (or other variant) expression with different dynamic ranges. We show its applicability by confirming known phenotypic effects for selected mutations for BRAF, HRAS, and SHP2.

Original language	English
Article number	17432
Number of pages	11
Journal	Scientific Reports
Volume	5
DOIs	https://doi.org/10.1038/srep17432
Publication status	Published - 27 Nov 2015

Keywords

Diseases
Proteins

Access to Document

10.1038/srep17432Licence: Unspecified

TEMTAC_2015_PDF
This work is licensed under a Creative Commons Attribution 4.0 International License.
Final published version, 982 KBLicence: CC BY

Handle.net

Persistent link

Professor Imre Berger
- imre.bergerbristol.acuk
- School of Biochemistry - Professor of Biochemistry
Person: Academic

Cite this

@article{c0293299a8d8419ba386e9fc53c58672,

title = "Tuneable endogenous mammalian target complementation via multiplexed plasmidbased recombineering",

abstract = "Understanding the quantitative functional consequences of human disease mutations requires silencing of endogenous genes and expression of mutants at close to physiological levels. Changing protein levels above or below these levels is also important for system perturbation and modelling. Fast design optimization demands flexible interchangeable cassettes for endogenous gene silencing and tuneable expression. Here, we introduce {\textquoteleft}TEMTAC{\textquoteright}, a multigene recombineering and delivery system for simultaneous siRNA-based knockdown and regulated mutant (or other variant) expression with different dynamic ranges. We show its applicability by confirming known phenotypic effects for selected mutations for BRAF, HRAS, and SHP2.",

keywords = "Diseases, Proteins",

author = "Violeta Beltran-Sastre and Hannah Benistry and Julia Burnier and Imre Berger and Luis Serrano and Christina Kiel",

year = "2015",

month = nov,

day = "27",

doi = "10.1038/srep17432",

language = "English",

volume = "5",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Springer Nature",

}

TY - JOUR

T1 - Tuneable endogenous mammalian target complementation via multiplexed plasmidbased recombineering

AU - Beltran-Sastre, Violeta

AU - Benistry, Hannah

AU - Burnier, Julia

AU - Berger, Imre

AU - Serrano, Luis

AU - Kiel, Christina

PY - 2015/11/27

Y1 - 2015/11/27

N2 - Understanding the quantitative functional consequences of human disease mutations requires silencing of endogenous genes and expression of mutants at close to physiological levels. Changing protein levels above or below these levels is also important for system perturbation and modelling. Fast design optimization demands flexible interchangeable cassettes for endogenous gene silencing and tuneable expression. Here, we introduce ‘TEMTAC’, a multigene recombineering and delivery system for simultaneous siRNA-based knockdown and regulated mutant (or other variant) expression with different dynamic ranges. We show its applicability by confirming known phenotypic effects for selected mutations for BRAF, HRAS, and SHP2.

AB - Understanding the quantitative functional consequences of human disease mutations requires silencing of endogenous genes and expression of mutants at close to physiological levels. Changing protein levels above or below these levels is also important for system perturbation and modelling. Fast design optimization demands flexible interchangeable cassettes for endogenous gene silencing and tuneable expression. Here, we introduce ‘TEMTAC’, a multigene recombineering and delivery system for simultaneous siRNA-based knockdown and regulated mutant (or other variant) expression with different dynamic ranges. We show its applicability by confirming known phenotypic effects for selected mutations for BRAF, HRAS, and SHP2.

KW - Diseases

KW - Proteins

U2 - 10.1038/srep17432

DO - 10.1038/srep17432

M3 - Article (Academic Journal)

C2 - 26612112

VL - 5

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 17432

ER -

Tuneable endogenous mammalian target complementation via multiplexed plasmidbased recombineering

Abstract

Keywords

Access to Document

Handle.net

Fingerprint

Profiles

Professor Imre Berger

Cite this