Teaching Cre to follow directions

Duyne, Gregory D. Van

doi:10.1073/pnas.0811624106

Cited by 6 publications

(5 citation statements)

References 9 publications

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“…We recently uncovered that SREBP-1 (sterol regulatory element-binding protein-1), a master transcription factor in the regulation of lipid metabolism (57-59), is highly upregulated in GBM and promotes fatty acid synthesis (49, 51, 53, 60-62). Moreover, we found that GBM cells take up large amounts of cholesterol through LDLR (low-density lipoprotein receptor) that is also upregulated by SREBP-1 (63).…”

Section: Lds and Glioblastomamentioning

confidence: 99%

Lipid droplets, potential biomarker and metabolic target in glioblastoma

Geng

Guo

2017

IMRJ

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Lipid droplets (LDs) are subcellular organelles that store large amounts of the neutral lipids, triglycerides (TG) and/or cholesteryl esters (CE). LDs are commonly formed in adipocytes, liver cells and macrophages, and their formation has been shown to be associated with the progression of metabolic diseases, i.e., obesity, fatty liver and atherosclerosis. Interestingly, LDs are also found in some tumor tissues. We recently showed that LDs are prevalent in glioblastoma (GBM), the most deadly brain tumor, but are not detectable in low-grade gliomas and normal brain tissues, suggesting that LDs may serve as a novel diagnostic biomarker for GBM. This short review will briefly introduce LD biology, summarize recent observations about LDs in several types of cancer tissues, and discuss LD formation in GBM. Moreover, we will highlight the role of SOAT1 (sterol-O transferase 1), a key enzyme regulating CE synthesis and LD formation in GBM, in the regulation of SREBP (sterol regulatory-element binding protein) activation. The therapeutic potential of LDs and SOAT1 will be discussed.

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Section: Lds and Glioblastomamentioning

confidence: 99%

Lipid droplets, potential biomarker and metabolic target in glioblastoma

Geng

Guo

2017

IMRJ

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“…Whereas neurophysiological and neuroimaging studies originally suggested that hyperexcitability and reorganization within the auditory cortex could explain audiological changes in tinnitus,(13) more recent work reveals involvement of two general domain networks:(14) a frontoparietal “working memory” network broadly involved in cognitive control (including the volitional control of attention)(15,16) and a cinguloinsular “saliency” network mediating attentional processing of pain and emotion. (17,18) In a corresponding fashion, rTMS treatments originally targeted temporal cortex to influence auditory processes, i.e.…”

Section: Introductionmentioning

confidence: 99%

“…STG has reciprocal connections to regions of the prefrontal cortex that mediate attention. To probe the hypothesized influence of STG stimulation on attention, a subset of patients with tinnitus enrolled in an rTMS clinical trial [n=12, 9 male, mean (sd) age = 49 (15) years] underwent an attentional conflict task before and after rTMS treatment in a repeated-measures functional magnetic resonance imaging (fMRI) study. …”

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confidence: 99%

Neural activity during attentional conflict predicts reduction in tinnitus perception following rTMS

et al. 2017

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Background Subjective idiopathic tinnitus is an intrusive, distracting, and potentially disabling disorder characterized by phantom perception of sounds. Although tinnitus has no approved pharmacologic treatment, recent evidence supports the use of repetitive transcranial magnetic stimulation (rTMS) to alleviate tinnitus symptoms. Objective/Hypothesis Repetitive TMS delivered over the middle superior temporal gyrus (STG) may alter ratings of tinnitus awareness and annoyance more than loudness due to change in attentional processing. STG has reciprocal connections to regions of the prefrontal cortex that mediate attention. To probe the hypothesized influence of STG stimulation on attention, a subset of patients with tinnitus enrolled in an rTMS clinical trial [n=12, 9 male, mean (sd) age = 49 (15) years] underwent an attentional conflict task before and after rTMS treatment in a repeated-measures functional magnetic resonance imaging (fMRI) study. Methods The Multi-Source Interference Task (MSIT), a Stroop-based visual attentional conflict fMRI task, was used to map participants’ neural processing of attentional conflict prior to rTMS intervention (Baseline) and after three rTMS intervention arms: Sham, 1Hz, and 10Hz (four sessions per arm, 1800 pulses per session, delivered @110% of the motor threshold over the posterior superior temporal gyrus). Results All measures of tinnitus severity (awareness, loudness, and annoyance) improved with 1Hz rTMS intervention; however, the greatest and most robust changes were observed for ratings of tinnitus awareness (mean 16% reduction in severity from Baseline, p<0.01). The MSIT elicited a similar pattern of neural activation among tinnitus participants at Baseline compared to an independent sample of 43 healthy comparison adults (r=0.801, p≤0.001). Linear regression with bootstrap resampling showed that greater recruitment of bilateral prefrontal and bilateral parietal regions by MSIT at Baseline corresponded with poorer treatment response. Individual regions’ activities explained 37–67% variance in participant treatment response, with left dorsolateral prefrontal cortex’s MSIT activity at Baseline explaining the greatest reduction in tinnitus awareness following 1 Hz stimulation. Although left dorsolateral prefrontal cortex activity at Baseline also predicted reduction in tinnitus loudness and annoyance (~50% variance explained), these symptoms were more strongly predicted by right middle occipital cortex (~70% variance explained) – suggesting that the neural predictors of symptom-specific treatment outcomes may be dissociable. Conclusion These candidate neural reactivity markers of treatment response have potential clinical value in identifying tinnitus sufferers who would or would not therapeutically benefit from rTMS intervention.

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“…These enzymes, including FLP recombinase [7], ΦC31 integrase [8–12], λ‐integrase [13], and Cre recombinase [14] facilitate genome editing functions such as deletions, insertions, inversions, and exchanges based on recognition of short, palindromic sequences that are typically integrated into the host's genome. Cre recombinase was prolifically utilized in early mouse recombineering efforts [15] and has several distinct advantages: it does not require protein factors [13], has high expression in mammalian systems [16], is more efficient than FLP recombinase [17] and ΦC31 integrase [18], and pseudo Cre recognition sites ( loxP ) do not interfere with recombinase activity in the mammalian genome [19]. Despite the fact that Cre recombinase is one of the most widely‐used tools for precise, site‐specific editing of mammalian genomes, its efficiency for transgene swapping and integration still remains limited [15, 20].…”

Section: Introductionmentioning

confidence: 99%

Using the Cre/loxsystem for targeted integration into the human genome:loxFAS‐loxP pairing and delayed introduction of Cre DNA improve gene swapping efficiency

Lanza

Dyess

Alper

2012

Biotechnology Journal

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Cre recombinase is a commonly-used genome editing tool suitable for site-specific integrations in mammalian genomes; however, the efficiency of transgenic swapping events compared to excision remains limited. Here we sought to identify important parameters and limiting factors that influence swapping propensity in this system, especially when using one wild-type loxP site. To modulate and increase the occurrence of swapping events, we identified two novel parameters. First, we identified the loxFAS-loxP pairing, a sequence never before used in mammalian systems, as the best choice for increasing swapping events in human cell lines. Second, for the first time we implicate the importance of delayed introduction of Cre DNA for optimal swapping efficiency. This same modification could potentially be of use to other systems catalyzing trimolecular reactions such as ΦC31 integrase and FLP recombinase where we hypothesize that transport of the exchange cassette is likewise initially rate limiting. The total number of recombination events, but not the ratio of swapping to excision, was found to be influenced by the quantity of Cre DNA transfected. Through this study, we were able to obtain Cre-mediated swapping frequencies of 8-12% without antibiotic enrichment, which represents nearly an order of magnitude increase over prior reports in the literature.

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Teaching Cre to follow directions

Cited by 6 publications

References 9 publications

Lipid droplets, potential biomarker and metabolic target in glioblastoma

Lipid droplets, potential biomarker and metabolic target in glioblastoma

Neural activity during attentional conflict predicts reduction in tinnitus perception following rTMS

Using the Cre/loxsystem for targeted integration into the human genome:loxFAS‐loxP pairing and delayed introduction of Cre DNA improve gene swapping efficiency

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