SINEUP Effector Domain

SINEUPTM Key characteristics

  • Upregulates proteins up to 10-fold
  • Simple mode of action
  • Rapid effect - works with transient transfection
  • No need to create stable cell lines
  • Very cost-effective
  • Target endogenous or exogenous genes
  • Target locus is unaffected
  • mRNA levels are unaltered
  • Works across a range of cell lines 

Use SINEUPTM to

  • Increase biomanufacturing productivity
  • Increase in-vitro translation yields
  • Validate microarray experiments
  • Manipulate cell phenotype
  • Screen new drug targets
  • Screen a gene across different cell lines
  • Screen genetic variants

Product Contents

pUC19-ED  SINEUPTM effector domain (1105bp) cloned into pUC19, 2 ug

User Guide

Brochure Protocol data

 

Description

Carrieri et al have shown that an antisense non-coding transcribed sequence (covering the start codon of a gene)  linked to an embedded repetitive SINEB2 element can activate polysomes for increased translation. 

This novel mechanism allows researchers to knockup (up-regulate) a specific gene simply via introduction of an Effector Domain (trans-regulatory element containing the SINEB2 element) fused to a gene specific Binding Domain (an antisense gene encompassing the start codon (ATG) sequence). Similar to RNAi, the transcription of the gene and the sequence of the gene is unaffected.

This SINEUPTM effector domain (ED) is exactly the same as reported by Carrieri et al. The domain is 1105 bp in length and has been sequence verified. It is provided cloned into pUC19 and contains a linker domain. It is ready for inserting your target gene-specific biding domain and subcloning into your desired expression vector.

 

SINEUP

 

AS Uchl1 is an antisense non-coding RNA containing a SINEB2 element with the 5’ overlapping the Uchl1 gene start codon.  Increased expression of AS Uchl1 from a transfected construct results in increased expression of Uchl1 protein without any increase in Uchl1 mRNA levels. Expression levels of Uchl1 protein can be titrated by modulating expression of a construct containing AS Uchl1.

Knock-Up your Protein

SINEUPTM non-coding RNA technology represents a breakthrough for the study and manipulation of eukaryotic genes. SINEUPTM allows up to 10-fold increases in expression levels of target proteins by increasing the efficiency of translation. It has been proposed that the mechanism is similar to IRES elements which increase the efficiency of translation initiation. Transcription of the target gene is unaltered. Similar to RNAi, which can be used to knockdown (down-regulate) protein levels, SINEUPTMtechnology does not rely on the direct manipulation of the target gene. 

Characterization of the SINEUPTM system, published in Nature (and unpublished data) have demonstrated increases in protein levels as high as 10-fold. mRNA levels are unaffected. SINEUPTM technology provides an additional layer of control over protein production which can be combined with existing tools.

SINEUPTM Technology

The cellular machinery that generates up-regulation of protein expression with SINEUPTM constructs has been shown to work for multiple genes and to be present in a range of mammalian cell types including murine and human cells. SINEUPTMwas developed following identification and characterization of the long non-coding RNA (lncRNA) AS Uchl1 in neuronal cells.

 

   
                             GFP

The ability of SINEUPTM constructs to increase protein expression is not restricted to Uchl1.  In HEK293 cells a SINEUPTM construct incorporating an antisense motif for GFP increases levels of GFP protein without any increase in GFP mRNA levels 

SINEUPTM can be combined with other genetic tools that act at the mRNA level to increase the overall amount of protein expression that can be achieved from a given cell type.

Note: SINEUPTM technology is licensed from TransSINE technologies. Commercial users require a license for all purposes, including research. Academic users do not require a license. For further information, please contact info@transsine.com

References

Long non-coding antisense RNA controls Uchl1 translation through an embedded SINEB2 repeat  Carrieri et al (2012) Nature 491:454-457. 

LncRNAs have a say in protein translation  Huarte (2012) Cell Research

Web coverage of SINEUPTM technology arond the World

Science Daily  Non-Coding Antisense RNA Can Be Used to Stimulate Protein Production

My Navi  (In Japanese)

Biotech (in Chinese)

Science System FVG (In Italian)

Science Research Blog (In Persian)

Blog Daum (in Korean)

News Medical (in Korean)

Effector domain Insert Sequence (1105 nucleotides) cloned into pUC19

The ED sequence (italics) extends from position 52 to 1156. 

TCTAGACTCG AGCGGCCGCC ACTGTGCTGG ATATCTGCAG AATTCGCCCT TCAGTGCTAG  

AGGAGGTCAG AAGAGGGCAT TGGATCCCCC AGAACTGGAG TTATACGGTA ACCTCGTGGT

GGTTGTGAAC CACCATGTGG ATGGATATTG AGTTCCAAAC ACTGGTCCTG TGCAAGAGCA

TCCAGTGCTC TTAAGTGCTG AGCCATCTCT TTAGCTCCAG TCTCTTAAAA AACAAACAAA

CGAACGAACA GCAAGGGAGC TGGGTATGAC AACACATACT ATAATTCTAG TACTCAGGAT

GCTGAAACAG GAGGATTGCC TGACTGGGAG ATATAAGGAG AATCTGTTGT CACCCCCACC

CCTCCCCATA AAGGCAGAAT AAAAGAACGT CCTATAAACA AATAAACAAA CAACCCAATA

AAACAAAACC AAGATCTCTC CACCTTTTCT TTGCTTTTTC AGACTTTGTA ATAAGGCCCT

TTGGAGTGCA GGATATTCGG CAGGACAAGC AGAGAGGGAG ACCATCAGTT CTTTCTTTGA

TCAAGAAGAC TATGTTCCTT AGCAAACTGG TGTGTATTAT CTCTTATGCA ATGAGCCTGG

AAAGAGGGCA CAGCCACCGA GGATGGTACA GCATGGATGG ATGGTACGCT ACAGAGACTC

GGGAGCCCAA CTGTGAGTGG CTGACTGGCA TGGTAGGTTC AGGGAAGAAT TGGCCTGTGA

AGAAAATGTT CTTGAAAAGT GAACAAGGTG CAGGAGGTAG GAGTGGGTCC TGGGCAAAGC

AGGGGGTGCA TCCCAGCCTC AGGGAATAGC ACAGCAGAGG TCTGTTGATG CATGCGAGTG

CATGACCTGC TTGCCAATAG ACGATCAAGA ATGGGCAAAG CATCATGGGT GATGAGTGGG

AGAGGGGATG AGACATTCCT TTCTCCCTGC TGAGACTTCC ATTGAACCGA TGAGTTCTGA

ATAGAAGATG CCCCCCCACC CCCCCACCAG TGTAGAATCT GAAGGGAGGC ATATATTACC

CTATATTACT CTGTGTTGGC GGCGAGCTAT CTGACAGCCA ACCTTCCCAT ACATTTCATT

GGGCATACAC TAATGACAGG AAGTTCCTTT TGCTTGTATG CAAGAGATGG CTCACACGAT

GGAGAATTTA ATCTTGAAGGGCGAATTCCACCACACTGGA CTAGTGGATC CGAGCTCGGT