Application of the BC1 RNA gene promoter for short hairpin RNA expression in cultured neuronal cells

Kobayashi, Shunsuke; Higuchi, Takashi; Anzai, Kentaro

doi:10.1016/j.bbrc.2005.07.033

Cited by 6 publications

(2 citation statements)

References 18 publications

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“…Small hairpin (shRNA) cassettes were constructed as previously described. 22 Two shRNA UNC5B-specific sequences were selected using the online Ambion design tool and cross referenced with the National Center for Biotechnology Information database (www.ncbi.nlm.nih.gov/ BLAST/) to confirm there was no cross-reactivity. Sequences were selected based on GenBank NM_170744 nucleotide sequences 3072-3090 and 571-589, respectively.…”

Section: Methodsmentioning

confidence: 99%

Axon guidance cue Netrin-1 has dual function in angiogenesis

Yang

Zou²,

Wang³

et al. 2007

Cancer Biology & Therapy

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Section: Methodsmentioning

confidence: 99%

Axon guidance cue Netrin-1 has dual function in angiogenesis

Yang

Zou²,

Wang³

et al. 2007

Cancer Biology & Therapy

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“…Notably, this portion corresponds to the tRNA portion of BC1 from which BC1 evolved 34 . While other studies have utilized this portion of BC1 with its endogenous upstream activating sequences for RNAi 28 …”

Section: Rnai and Minimal Toxicity By Bc1(id)mentioning

confidence: 99%

The Human L1 Element Causes DNA Double-Strand Breaks in Breast Cancer

Deininger¹

2006

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Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Department of Defense, Washington Headquarters Services, Directorate for Information Operations and Reports (0704-0188), 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302. Respondents should be aware that notwithstanding any other provision of law, no person shall be subject to any penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. (From -To) REPORT DATE (DD-MM-YYYY) 01-08-2006 REPORT TYPE Final DATES COVERED PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) 8. PERFORMING ORGANIZATION REPORT NUMBERTulane University Health Science Center New Orleans, LA 70112 SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSOR/MONITOR'S ACRONYM(S) U.S. Army Medical Research and Materiel Command Fort Detrick, Maryland 21702-5012 SPONSOR/MONITOR'S REPORT NUMBER(S) DISTRIBUTION / AVAILABILITY STATEMENTApproved for Public Release; Distribution Unlimited SUPPLEMENTARY NOTES ABSTRACTThe source of mutagenesis for the establishment and maintenance of cancer is complex. However, defects in DNA repair genes in the double-strand break repair pathway are cancer predisposing. My lab has characterized a new potentially important source of double-strand breaks (DSBs) in human cells and are interested in characterizing which DNA repair genes act on this particular source of DNA damage. Selfish DNA accounts for 45% of the human genome. We have recently demonstrated that one particular selfish DNA, the L1 retrotransposon, creates DSBs via its endonuclease domain. The important conclusions from the published work are that 1) the human retrotransposon L1 creates DSBs and 2) the DSB repair gene ATM is required for L1 retrotransposition.To additionally characterize the roles of ATM and the ATM-related genes, BRCA1 and BRCA2, in L1-induced DSBs, we developed a new vector system to suppress their expression which is compatible with L1 assays. These constructs should also be of general utility. SUBJECT TERMSBRCA1, BRCA2, double-strand breaks, retrotransposition, LINE-1, ATM IntroductionThe source of mutagenesis for the establishment and maintenance of cancer is complex. However, defects in DNA repair genes in the double-strand break repair pathway are cancer predisposing. My lab has characterized a new potentially important source of double-strand breaks (DSBs) in human cells and are interested in characterizing which DNA repair genes act on this particular source of DNA damage. Prior to the funding of this grant we had unpublished data demonstrating that the human...

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Short Hairpin RNA-Mediated Gene Silencing

Lambeth

Smith

2012

Methods in Molecular Biology

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Since the first application of RNA interference (RNAi) in mammalian cells, the expression of short hairpin RNAs (shRNAs) for targeted gene silencing has become a benchmark technology. Using plasmid and viral vectoring systems, the transcription of shRNA precursors that are effectively processed by the RNAi pathway can lead to potent gene knockdown. The past decade has seen continual advancement and improvement to the various strategies that can be used for shRNA delivery, and the use of shRNAs for clinical applications is well underway. Driving these developments has been the many benefits afforded by shRNA technologies, including the stable integration of expression constructs for long-term expression, infection of difficult-to-target cell lines and tissues using viral vectors, and the temporal control of shRNA transcription by inducible promoters. The use of different effector molecule formats, promoters, and vector types, has meant that experiments can be tailored to target specific cell types and minimize cellular toxicities. Through the application of combinatorial RNAi (co-RNAi), multiple shRNA delivery strategies can improve gene knockdown, permit multiple transcripts to be targeted simultaneously, and curtail the emergence of viral escape mutants. This chapter reviews the history, cellular processing, and various applications of shRNAs in mammalian systems, including options for effector molecule design, vector and promoter types, and methods for multiple shRNA delivery.

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Application of the BC1 RNA gene promoter for short hairpin RNA expression in cultured neuronal cells

Cited by 6 publications

References 18 publications

Axon guidance cue Netrin-1 has dual function in angiogenesis

Axon guidance cue Netrin-1 has dual function in angiogenesis

The Human L1 Element Causes DNA Double-Strand Breaks in Breast Cancer

Short Hairpin RNA-Mediated Gene Silencing

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