ברצוננו להודות התרומות של כל עמיתיו ושותפיו ללימודים המקורי שלנו, אשר התוצאות כבר התייחסו בכתב היד (להמחשה). העבודה של המחברים נתמך על ידי מענקים הלאומית המכון של שיניים, מחקר ואסטתיים של מכוני הבריאות הלאומיים (1R01DE026043) ל AB ו מן האוניברסיטה של מחלת California Tobacco-Related מחקר לתוכנית AB ( TRDRP-26IR-0015) סטריט (TRDRP-25IP-0001). נותני החסות של המחקר שהיה אין תפקיד תכנון המחקר, איסוף נתונים, ניתוח נתונים, נתונים לפרשנות, כתיבת הדו ח, או ההחלטה להגיש לפרסום.

כל המחברים מצהירים ללא ניגוד אינטרסים.

Λ cII בחירה וזמינותו משמש לצורך זיהוי של מוטציות transgene cII התאוששה הדנ א הגנומי של תאים שמקורם איברים/רקמות של BB מכרסמים3. הגנום של בעלי חיים אלה הטרנסגניים מכילה עותקים של וקטור הסעות λLIZ chromosomally משולב, אשר נושאת את cII טנדם (294 bp) ו לצי (1,080 bp) transgenes, כמו גנים כתב mutati...

מגוון רחב של מודלים בבעלי חיים מהונדס ומערכות איתור מוטציה פותחו לבדיקת המוטגניות של חומרים מסרטנים בתאי יונקים. אלה, העכברים הטרנסגניים Big Blue (המכונים להלן BB), בחר λ cII מערכת איתור מוטציה יש כבר מועסקים לניסויים המוטגניות על ידי קבוצה זו רבים אחרים מחקר קבוצות ברחבי העולם1,2, 3,4,5,6,7,8,9. במשך 16 השנים האחרונות, חקרנו את ההשפעות המוטגניות של סוכנים כימי ו/או גופניים שונים באמצעות בעלי חיים אלה הטרנסגניים או שלהם תרביות תאים עובריים פיברובלסט המקביל שטופלו תרכובת המבחן ולאחר לאחר מכן ניתח גן # מושגים בסיסיים, גנוטיפ של transgene cII מאת λ cII בחירה וזמינותו של רצפי DNA, בהתאמה10,11,12,13,14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24. הגנום של בעלי חיים אלה הטרנסגניים מכיל bacteriophage λ הסעות וקטור (λLIZ) משולב על כרומוזום 4 כמו2,1,concatemer ראש זנב מרובת עותקים25. הווקטור הסעות λLIZ נושאת שני גנים כתב mutational, כלומר לצי של cII transgenes1,2,25,26,27, 28,29,30,31,32,33,34,35,36, 37,38,39,40,41,42,43,44,45 , 46 , 47. λ cII בחירה וזמינותו מבוססת על החזרתה של הווקטורים הסעות λLIZ של ה-DNA גנומי של תאים שמקורם איברים/רקמות חיות הטרנסגניים1,2,25 . הווקטורים הסעות λLIZ התאושש ארוזים ואז לתוך ראשי phage λ מסוגל להדביק מארח מחוון Escherichia coli. לאחר מכן, נגוע החיידקים גדלים בתנאים סלקטיבית כדי לאפשר את הניקוד וניתוח של מוטציות של cII transgene1,3.
כאן, אנו מתארים את פרוטוקול מפורט עבור וזמינותו בחר cII λ, אשר מורכב של בידוד של דנ א גנומי של תאים/איברים של חיות הטרנסגניים מטופלים במבחנה/ויוו עם החזרה במתחם, בדיקה של λLIZ מעבורת וקטורים מן ה-DNA גנומי, אריזה של הווקטורים לתוך phages λ זיהומיות, זיהום של החיידק המארח עם bacteriophages, זיהוי cII-המוטציות בתנאים סלקטיבית כדי לקבוע את תדירות מוטציה של cII , ו ניתוח רצף הדנ א להקים cII מוטציה הספקטרום. הפרוטוקול ניתן להחיל על העכבר הטרנסגניים/חולדה תא תרבויות מטופלים במבחנה עם סוכן כימי/פיזי של עניין, או רקמות/איברים מבעלי החיים המתאים שטופל ויוו עם בדיקה כימית/סוכן1, 2,4,48,49,50,51,52. מצגת סכמטי של λ cII בחירה וזמינותו מוצג באיור1.

<table><tbody><tr><td>Agar</td><td>MO Bio Laboratories, Inc.</td><td>12112-05</td><td>Bacteriological grade</td></tr><tr><td>BigDye Terminator v3.1 Cycle Sequencing Kit</td><td>Thermo Fisher Scientific</td><td>4337455</td><td>None</td></tr><tr><td>Casein Peptone</td><td>Alfa Aesar</td><td>H26557</td><td>None</td></tr><tr><td>Gelatine</td><td>J. T. Baker</td><td>2124-01</td><td>Powder</td></tr><tr><td>Glycerol</td><td>Fisher Scientific</td><td>BP 229-1 / M-13750</td><td>None</td></tr><tr><td>LB Agar</td><td>Fisher Scientific</td><td>BP 9724-500</td><td>None</td></tr><tr><td>QIAquick PCR purification kit</td><td>Qiagen</td><td>8104</td><td>50 PCR purification reactions</td></tr><tr><td>Sodium Acetate Trihydrate</td><td>Fisher Scientific</td><td>M-15756</td><td>None</td></tr><tr><td>&lt;em&gt;Taq5000&lt;/em&gt; DNA Polymerase&amp;nbsp;</td><td>Qiagen</td><td>201207</td><td>None</td></tr><tr><td>Thiamine Hydrochloride</td><td>Macron Fine Chemicals</td><td>2722-57</td><td>None</td></tr><tr><td>Transpack Packaging Extract</td><td>Stratagene Corp., Acquired by BioReliance | Sigma-Aldrich Corp.</td><td>200223</td><td>50 packaging reactions</td></tr><tr><td>Tris Base</td><td>Fisher Scientific</td><td>BP 152-1 / EC 201-064-4</td><td>None</td></tr><tr><td>Trypton</td><td>Biosciences</td><td>RC-110</td><td>None</td></tr></tbody></table>

the lambda select cii mutation detection system

מספר מודלים בבעלי חיים מהונדס, מערכות זיהוי המוטציה פותחו לבדיקת המוטגניות של חומרים מסרטנים בתאי יונקים. אלה, העכברים הטרנסגניים, בחר למבדה (λ) cII מערכת איתור מוטציה יש כבר מועסקים לניסויים המוטגניות על ידי קבוצות מחקר רבות ברחבי העולם. כאן, אנו מתארים את פרוטוקול מפורט עבור למדא בחר cII מוטציה וזמינותו, אשר ניתן להחיל על תאים בתרבית של העכברים הטרנסגניים/חולדות או החיות המקביל שטופלו עם סוכן כימי/פיזי של עניין. הפרוטוקול מורכב מהשלבים הבאים: (1) בידוד של דנ א גנומי מתאי או איברים/רקמות חיות הטרנסגניים שטופלו חוץ גופית או ויוו, בהתאמה, תרכובת מבחן; (2) שחזור של וקטור הסעות למדא נושא גן כתב mutational (כלומר, cII transgene) של ה-DNA גנומי; (3) אריזה של הווקטורים שניצלו לתוך bacteriophages זיהומיות; (4) להדביק חיידקים המארח ו culturing בתנאים סלקטיבית כדי לאפשר הפצת מוטציות המושרה cII ; (5) הקלפים של cII-מוטציות ו- DNA רצף ניתוח כדי לקבוע את תדירות מוטציה cII ואת הספקטרום מוטציה, בהתאמה.

בהתאם לנתונים הפצה, בדיקות פרמטרית או שאינם פרמטרי משמשים כדי לקבוע את המשמעות של הבדל בשכיחות מוטציה cII בין קבוצות הטיפול ושליטה (קרי, המושרה לעומת תדרים מוטציה ספונטנית) . השוואה של התדרים מוטציה המושרה cII קבוצות טיפול שונה נעשית על ידי שונים (pairwise) בדיקות ?...

Watch this Scientific Journal Video about The Lambda Select cII Mutation Detection System at JoVE.com

The Lambda Select cII Mutation Detection System

למדא בחר cII מערכת איתור מוטציה

אנו מתארים פרוטוקול מפורט את assay מוטציה cII למדא בחר בתאים בתרבית של מכרסמים הטרנסגניים או החיות המקביל שטופלו עם סוכן כימי/פיזי של עניין. גישה זו כבר בשימוש נרחב לניסויים המוטגניות של חומרים מסרטנים בתאי יונקים.

A number of transgenic animal models and mutation detection systems have been developed for mutagenicity testing of carcinogens in mammalian cells. Of ...

the-lambda-select-cii-mutation-detection-system

Research

JoVE Journal

Immunology and Infection

The Lambda Select cII Mutation Detection System

7.8K Views.  University of Southern California. אנו מתארים פרוטוקול מפורט את assay מוטציה cII למדא בחר בתאים בתרבית של מכרסמים הטרנסגניים או החיות המקביל שטופלו עם סוכן כימי/פיזי של עניין. גישה זו כבר בשימוש נרחב לניסויים המוטגניות של חומרים מסרטנים בתאי יונקים.

Video: The Lambda Select cII Mutation Detection System

Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors

As our understanding of the driver mutations necessary for initiation and progression of cancers improves, we gain critical information on how specific molecular profiles of a tumor may predict responsiveness to therapeutic agents or provide knowledge about prognosis. At our institution a tumor genotyping program was established as part of routine clinical care, screening both hematologic and solid tumors for a wide spectrum of mutations using two next-generation sequencing (NGS) panels: a custom, 33 gene hematological malignancies panel for use with peripheral blood and bone marrow, and a commercially produced solid tumor panel for use with formalin-fixed paraffin-embedded tissue that targets 47 genes commonly mutated in cancer. Our workflow includes a pathologist review of the biopsy to ensure there is adequate amount of tumor for the assay followed by customized DNA extraction is performed on the specimen. Quality control of the specimen includes steps for quantity, quality and integrity and only after the extracted DNA passes these metrics an amplicon library is generated and sequenced. The resulting data is analyzed through an in-house bioinformatics pipeline and the variants are reviewed and interpreted for pathogenicity. Here we provide a snapshot of the utility of each panel using two clinical cases to provide insight into how a well-designed NGS workflow can contribute to optimizing clinical outcomes.

This manuscript describes clinical protocols for two next-generation sequencing panels. One panel interrogates hematologic malignancies while the other panel targets genes commonly mutated in solid tumors. Molecular classification of driver mutations in human malignancies offers valuable prognostic and predictive information.

רצף הדור הבא לצורך זיהוי של מוטציות שניתן לפעול ב מוצק לנוזל גידולים

As our understanding of the driver mutations necessary for initiation and progression of cancers improves, we gain critical information on how specific ...

Cancer Research

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Accurate detection and identification of low frequency mutations can be problematic when assessing residual disease after therapy, screening for emerging resistance mutations during therapy, or when patients have few circulating tumor cells. Wild-type blocking PCR followed by sequencing analysis offers high sensitivity, flexibility, and simplicity as a methodology for detecting these low frequency mutations. By adding a custom designed locked nucleic acid oligonucleotide to a new or previously established conventional PCR based sequencing assay, sensitivities of approximately 1 mutant allele in a background of 1,000 WT alleles can be achieved (1:1,000). Sequencing artifacts associated with deamination events commonly found in formalin fixed paraffin embedded tissues can be partially remedied by the use of uracil DNA glycosylase during extraction steps. The optimized protocol here is specific for detecting MYD88 mutation, but can serve as a template to design any WTB-PCR assay. Advantages of the WTB-PCR assay over other commonly utilized assays for the detection of low frequency mutations including allele specific PCR and real-time quantitative PCR include fewer occurrences of false positives, greater flexibility and ease of implementation, and the ability to detect both known and unknown mutations.

Wild-type blocking PCR followed by direct sequencing offers a highly sensitive method of detection for low frequency somatic mutations in a variety of sample types.

חסימת PCR משולבת מסוג Wild עם רצף ישיר כשיטה רגישה עבור איתור של תדרים נמוכים מוטציות סומטיות

Accurate detection and identification of low frequency mutations can be problematic when assessing residual disease after therapy, screening for emerging ...

Genetics

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing

The analysis of circulating tumor DNA (ctDNA) using next-generation sequencing (NGS) has become a valuable tool for the development of clinical oncology. However, the application of this method is challenging due to its low sensitivity in analyzing the trace amount of ctDNA in the blood. Furthermore, the method may generate false positive and negative results from this sequencing and subsequent analysis. To improve the feasibility and reliability of ctDNA detection in the clinic, here we present a technique which enriches rare mutations for sequencing, Enrich Rare Mutation Sequencing (ER-Seq). ER-Seq can distinguish a single mutation out of 1 x 107 wild-type nucleotides, which makes it a promising tool to detect extremely low frequency genetic alterations and thus will be very useful in studying disease heterogenicity. By virtue of the unique sequencing adapter&#39;s ligation, this method enables an efficient recovery of ctDNA molecules, while at the same time correcting for errors bidirectionally (sense and antisense). Our selection of 1021 kb probes enriches the measurement of target regions that cover over 95% of the tumor-related driver mutations in 12 tumors. This cost-effective and universal method enables a uniquely successful accumulation of genetic data. After efficiently filtering out background error, ER-seq can precisely detect rare mutations. Using a case study, we present a detailed protocol demonstrating probe design, library construction, and target DNA capture methodologies, while also including the data analysis workflow. The process to carry out this method typically takes 1-2 days.

This manuscript describes a technique for detecting mutations of low frequency in ctDNA, ER-Seq. This method is differentiated by its unique use of two-directional error correction, a special background filter, and efficient molecular acquirement.

זיהוי של מוטציות נדירות CtDNA באמצעות רצף הדור הבא

The analysis of circulating tumor DNA (ctDNA) using next-generation sequencing (NGS) has become a valuable tool for the development of clinical oncology. ...

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Conventional next-generation sequencing techniques (NGS) have allowed for immense genomic characterization for over a decade. Specifically, NGS has been used to analyze the spectrum of clonal mutations in malignancy. Though far more efficient than traditional Sanger methods, NGS struggles with identifying rare clonal and subclonal mutations due to its high error rate of ~0.5&#8211;2.0%. Thus, standard NGS has a limit of detection for mutations that are &#62;0.02 variant allele fraction (VAF). While the clinical significance for mutations this rare in patients without known disease remains unclear, patients treated for leukemia have significantly improved outcomes when residual disease is &#60;0.0001 by flow cytometry. In order to mitigate this artefactual background of NGS, numerous methods have been developed. Here we describe a method for Error-corrected DNA and RNA Sequencing (ECS), which involves tagging individual molecules with both a 16 bp random index for error-correction and an 8 bp patient-specific index for multiplexing. Our method can detect and track clonal mutations at variant allele fractions (VAFs) two orders of magnitude lower than the detection limit of NGS and as rare as 0.0001 VAF.

Next-generation sequencing (NGS) is a powerful tool for genomic characterization that is limited by the high error rate of the platform (~0.5&#8211;2.0%). We describe our methods of error-corrected sequencing that allow us to obviate the NGS error rate and detect mutations at variant allele fractions as rare as 0.0001.

אירוע נדיר איתור שגיאות DNA ו RNA רצף

Conventional next-generation sequencing techniques (NGS) have allowed for immense genomic characterization for over a decade. Specifically, NGS has been ...

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Droplet digital polymerase chain reaction (ddPCR) is a highly sensitive quantitative polymerase chain reaction (PCR) method based on sample fractionation into thousands of nano-sized water-in-oil individual reactions. Recently, ddPCR has become one of the most accurate and sensitive tools for circulating tumor DNA (ctDNA) detection. One of the major limitations of the standard ddPCR technique is the restricted number of mutations that can be screened per reaction, as specific hydrolysis probes recognizing each possible allelic version are required. An alternative methodology, the drop-off ddPCR, increases throughput, since it requires only a single pair of probes to detect and quantify potentially all genetic alterations in the targeted region. Drop-off ddPCR displays comparable sensitivity to conventional ddPCR assays with the advantage of detecting a greater number of mutations in a single reaction. It is cost-effective, conserves precious sample material, and can also be used as a discovery tool when mutations are not known a priori.

Here, we present a protocol to accurately quantify multiple genetic alterations of a target region in a single reaction using drop-off ddPCR and a unique pair of hydrolysis probes.

Droplet יחיד שרשרת התגובה פולימראז דיגיטלי עבור זיהוי מקיף, בו זמנית של מוטציות באזורים חמה

Droplet digital polymerase chain reaction (ddPCR) is a highly sensitive quantitative polymerase chain reaction (PCR) method based on sample fractionation ...

Efficient Purification and LC-MS/MS-based Assay Development for Ten-Eleven Translocation-2 5-Methylcytosine Dioxygenase

The epigenetic transcription regulation mediated by 5-methylcytosine (5mC) has played a critical role in eukaryotic development. Demethylation of these epigenetic marks is accomplished by sequential oxidation by ten-eleven translocation dioxygenases (TET1-3), followed by the thymine-DNA glycosylase-dependent base excision repair. Inactivation of the TET2 gene due to genetic mutations or by other epigenetic mechanisms is associated with a poor prognosis in patients with diverse cancers, especially hematopoietic malignancies. Here, we describe an efficient single step purification of enzymatically active untagged human TET2 dioxygenase using cation exchange chromatography. We further provide a liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach that can separate and quantify the four normal DNA bases (A, T, G, and C), as well as the four modified cytosine bases (5-methyl, 5-hydroxymethyl, 5-formyl, and 5-carboxyl). This assay can be used to evaluate the activity of wild type and mutant TET2 dioxygenases.

Here, we present a protocol for an efficient single step purification of the active untagged human ten-eleven translocation-2 (TET2) 5-methylcytosine dioxygenase using ion-exchange chromatography and its assay using a liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based approach.

טיהור יעיל ופיתוח Assay LC-MS/MS-מבוסס על 10-11 טרנסלוקציה-2 5-Methylcytosine Dioxygenase

The epigenetic transcription regulation mediated by 5-methylcytosine (5mC) has played a critical role in eukaryotic development. Demethylation of these ...

Biochemistry

qKAT: Quantitative Semi-automated Typing of Killer-cell Immunoglobulin-like Receptor Genes

Killer cell immunoglobulin-like receptors (KIRs) are a set of inhibitory and activating immune receptors, on natural killer (NK) and T cells, encoded by a polymorphic cluster of genes on chromosome 19. Their best-characterized ligands are the human leukocyte antigen (HLA) molecules that are encoded within the major histocompatibility complex (MHC) locus on chromosome 6. There is substantial evidence that they play a significant role in immunity, reproduction, and transplantation, making it crucial to have techniques that can accurately genotype them. However, high-sequence homology, as well as allelic and copy number variation, make it difficult to design methods that can accurately and efficiently genotype all KIR genes. Traditional methods are usually limited in the resolution of data obtained, throughput, cost-effectiveness, and the time taken for setting up and running the experiments. We describe a method called quantitative KIR semi-automated typing (qKAT), which is a high-throughput multiplex real-time polymerase chain reaction method that can determine the gene copy numbers for all genes in the KIR locus. qKAT is a simple high-throughput method that can provide high-resolution KIR copy number data, which can be further used to infer the variations in the structurally polymorphic haplotypes that encompass them. This copy number and haplotype data can be beneficial for studies on large-scale disease associations, population genetics, as well as investigations on expression and functional interactions between KIR and HLA.

Quantitative killer cell immunoglobulin-like receptor (KIR) semi-automated typing (qKAT) is a simple, high-throughput, and cost-effective method to copy number type KIR genes for their application in population and disease association studies.

qKAT: Semi-automated כמותיים הקלדה של הרוצח-תא גנים קולטני דמוי נוגדן

Killer cell immunoglobulin-like receptors (KIRs) are a set of inhibitory and activating immune receptors, on natural killer (NK) and T cells, encoded by a ...

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Complications associated with upfront and repeat surgical tissue sampling present the need for minimally invasive platforms capable of molecular sub-classification and temporal monitoring of tumor response to therapy. Here, we describe our dPCR-based method for the detection of tumor somatic mutations in cell free DNA (cfDNA), readily available in&#160;patient biofluids. Although limited in the number of mutations that can be tested for in each assay, this method provides a high level of sensitivity and specificity. Monitoring of mutation abundance, as calculated by MAF, allows for the evaluation of tumor response to therapy, thereby providing a much-needed supplement to radiographic imaging.

Here, we present a protocol to detect tumor somatic mutations in circulating DNA present in patient biological fluids (biofluids). Our droplet digital polymerase chain reaction (dPCR)-based method enables quantification of the tumor mutation allelic frequency (MAF), facilitating a minimally invasive complement to diagnosis and temporal monitoring of tumor response.

איתור וניטור של DNA הקשורים במחזור הגידול ב ביולואידים חולה

Complications associated with upfront and repeat surgical tissue sampling present the need for minimally invasive platforms capable of molecular ...

Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells

In recent years, it has become apparent that genomic instability is tightly related to many developmental disorders, cancers, and aging. Given that stem cells are responsible for ensuring tissue homeostasis and repair throughout life, it is reasonable to hypothesize that the stem cell population is critical for preserving genomic integrity of tissues. Therefore, significant interest has arisen in assessing the impact of endogenous and environmental factors on genomic integrity in stem cells and their progeny, aiming to understand the etiology of stem-cell based diseases.
LacI transgenic mice carry a recoverable &#955;&#160;phage vector encoding the LacI reporter system, in which the LacI gene serves as the mutation reporter. The result of a mutated LacI gene is the production of &#946;-galactosidase that cleaves a chromogenic substrate, turning it blue. The LacI reporter system is carried in all cells, including stem/progenitor cells and can easily be recovered and used to subsequently infect E. coli. After incubating infected E. coli on agarose that contains the correct substrate, plaques can be scored; blue plaques indicate a mutant LacI gene, while clear plaques harbor wild-type. The frequency of blue (among clear) plaques indicates the mutant frequency in the original cell population the DNA was extracted from. Sequencing the mutant LacI gene will show the location of the mutations in the gene and the type of mutation.
The LacI transgenic mouse model is well-established as an in vivo mutagenesis assay. Moreover, the mice and the reagents for the assay are commercially available. Here we describe in detail how this model can be adapted to measure the frequency of spontaneously occurring DNA mutants in stem cell-enriched Lin-IL7R-Sca-1+cKit++(LSK) cells and other subpopulations of the hematopoietic system.

Here we describe an in vivo mutagenesis assay for small numbers of purified hematopoietic cells using the LacI transgenic mouse model.&#160;The LacI gene can be isolated to determine the frequency, location, and type of DNA mutants&#160;spontaneously arisen or after exposure to genotoxins.

זיהוי מוטציות DNA בתאי גזע / אב המטוהר Hematopoietic

In recent years, it has become apparent that genomic instability is tightly related to many developmental disorders, cancers, and aging. Given that stem ...

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

Genome-scale interrogation of gene function using RNA interference (RNAi) holds tremendous promise for the rapid identification of chemically tractable cancer cell vulnerabilities. Limiting the potential of this technology is the inability to rapidly delineate the mechanistic basis of phenotypic outcomes and thus inform the development of molecularly targeted therapeutic strategies. We outline here methods to deconstruct cellular phenotypes induced by RNAi-mediated gene targeting using multiplexed reporter systems that allow monitoring of key cancer cell-associated processes. This high-content screening methodology is versatile and can be readily adapted for the screening of other types of large molecular libraries.

Achieving a systems level understanding of cellular processes is a goal of modern-day cell biology. We describe here strategies for multiplexing luciferase reporters of various cellular function end-points to interrogate gene function using genome-scale RNAi libraries.

פלטפורמת הקרנה מבוססת לוציפראז Multiplexed לחקירת מלחמה בסרטן הקשורים להעברת אותות בתאים בתרבית

Genome-scale interrogation of gene function using RNA interference (RNAi)  holds tremendous promise for the rapid identification of chemically tractable  ...