WEB BASED VECTOR DESIGN

FIELD OF THE INVENTION

The invention relates generally to computer methods and systems for acquiring, storing, manipulating, analyzing, displaying and interacting with information relating to vector design and construction, for linking such elements of vector design to other information, such as information in RNAi databases, and for storing, retrieving, manipulating, analyzing, displaying, and interacting with the linked information for purposes of designing and purchasing a virus vector.

BACKGROUND OF THE INVENTION

Computational methods and systems are essential in virtually every area of human endeavor. Often, the limits of a computational system define abilities to acquire, manipulate, analyze, understand or utilize information. This effect is becoming increasingly important in many areas of biochemical research, such as genomics research and research on gene expression. Many computational methods and systems have been developed in these areas and help researchers identify a gene, or genes, implicated in a specific disease and have lead to the development of innovative therapies.

Gene therapy is a scientific discipline devoted to the development of therapies wherein one or more genes, anti-sense, RNAi, or an equivalent agent, (i.e., "biomolecule") is delivered to a mammal by way of a vector. When a biomolecule and vector are designed correctly and introduced into the mammal, the vector construct has been shown to alleviate disease. Many vectors are available to enable the delivery of biomolecules such as plasmids, adenovirus, retrovirus, and lentivirus. However, the expert in the specific therapeutic field such as oncology, diabetes, or asthma, who has identified a biomolecule able to alleviate the affects of disease, is not an expert in the field of virology and designing a vector for delivering one or more biomolecules into a mammal. Therefore, there is a tremendous need to provide computational methods and systems to facilitate design of vector constructs, that is, the combination of a virus vector and a biomolecule.

SUMMARY OF THE INVENTION

Accordingly, it is an object of the present invention to meet the foregoing needs by providing methods and systems enabling the design of vector constructs and that allow researchers to easily design and/or purchase one or more vector(s) on-line using a web based system.

One embodiment of the present invention is a method in a computer system for generating a graphic display for visualization of one or more vector construct(s). This method comprises: providing an area for the insertion of a nucleic acid sequence of a biomolecule; placing the nucleic acid sequence of a biomolecule into the area; inserting the nucleic acid sequence of a biomolecule into a vector, wherein the placement of the nucleic acid sequence of a biomolecule into the vector creates a theoretical functional vector construct; and purchasing the theoretical functional vector construct on the graphic display. The vectors used in the methods of the present invention may be a plasmid, virus, or other construct, but preferably is a lentivirus. The biomolecule used in the methods of the present invention is a genetic element including, but not limited to: a gene, RNAi, promoter, enhancer element, reporter gene, selection gene, envelope protein, or any combination thereof. It is also preferred that the methods of the present invention generate a graphic display wherein a click agreement is viewed by an operator of the computer system prior to purchasing the theoretical functional vector. Another embodiment of the present invention is a method in a computer system for generating a graphic display for visualization of one or more vector construct(s), comprising: providing a first area for the insertion of a nucleic acid sequence of a gene; placing the nucleic acid sequence of a gene into the first area; inserting the nucleic acid sequence of a gene into a vector, wherein the placement of the nucleic acid sequence of a gene into the vector creates a theoretical functional vector construct; and purchasing the theoretical functional vector construct observed on the graphic display. This method may comprise a second area or more, for the insertion of one or more biomolecules into the vector.

Another embodiment of the present invention is a method in a computer system for generating a graphic display for visualization of one or more vector construct(s), comprising: providing a first area for the insertion of a nucleic acid sequence of a RNAi; placing the nucleic acid sequence of a RNAi into the first area; inserting the nucleic acid sequence of a RNAi into a vector, wherein the placement of the nucleic acid sequence of a RNAi into the vector creates a theoretical functional vector construct; and purchasing the theoretical functional vector construct on the graphic display. It is preferable that the RNAi

is a shRNA or a micro RNA. This methods of the present invention may comprise a second area, or more (such as a third area, fourth area, fifth area, etc.), for the insertion of one or more biomolecules (the first biomolecule in the first area, the second biomolecule into the second area, the third biomolecule into the third area, etc., or any combination thereof).

BRIEF DESCRIPTION OF THE INVENTION

FIG. 1 Starting Page For Designing a Custom Vector

FIG. 2 First Tab: For Designing a Custom Vector Containing a Gene FIG. 3 Second Tab: For Designing a Custom Vector Containing a Gene

FIG. 4 Third Tab: For Designing a Custom Vector Containing a Gene

FIG. 5 Fourth Tab: For Designing a Custom Vector Containing a Gene

FIG. 6 Fifth Tab: For Designing a Custom Vector Containing a Gene

FIG. 7 Sixth Tab: Review a Vector FIG. 8 Seventh Tab: Checkout Screen

FIG. 9 Seventh Tab: Checkout Screen

FIG. 10 First Tab: For Designing a Custom Vector Containing a RNAi

FIG. 11 Second Tab: For Designing a Custom Vector Containing a RNAi

FIG. 12 Third Tab: For Designing a Custom Vector Containing a RNAi FIG. 13 Fourth Tab: For Designing a Custom Vector Containing a RNAi

FIG. 14 Fifth Tab: For Designing a Custom Vector Containing a RNAi

FIG. 15 Sixth Tab: For Designing a Custom Vector Containing a RNAi

FIG. 16 Seventh Tab: Review a Vector

DETAILED DESCRIPTION OF THE INVENTION

Reference will now be made to preferred embodiments of this invention, examples of which will be obvious from the detailed description of the invention. The present invention relates to methods and systems enabling the design of vector constructs and that allow researchers to easily design and/or purchase vector(s) on-line using a web based system. In order to better understand the invention the following terms have been defined:

"biomolecule" shall mean a genetic element, including but not limited to, a gene, RNAi, promoter, enhancer element, reporter gene, selection gene, envelope protein.

"click agreement" shall mean a document explaining the terms and conditions of the sale of a theoretically functional vector.

"gene silencing sequence" shall mean the nucleic acid sequence of an RNAi.

"lentiviral" shall mean any vector derived from a lentivirus.

"reporter gene" shall mean some gene, or sequence, that produces a protein that is capable of being assayed and/or quantified.

"RNAi" shall mean any RNA that inhibits a gene including, but not limited to, shRNAi, miRNA, and siRNA.

"screen" shall mean an area that can be viewed on a computer.

"selection gene" shall mean any gene or sequence that allows for the selection of cells including, but not limited to, neomycin and puromycin.

"theoretical functional vector" shall mean a vector on a graphic display having all the genetic elements required to express a biomolecule in a cell.

"turbo-GFP" shall mean a variant of GFP (i.e. green fluorescent protein)

"vector" shall mean any genetic delivery system including, but not limited to, plasmids, virus, bacteria, protein, lipid, polymer, and chemical.

Graphic Display

The graphic display of the methods and systems of the present invention are illustrated, but not limited to, the graphic displays shown in FIGS. 1-16. A user may log on to a host's web site and be prompted to design a vector. Specifically, the user may be asked if he or she would like to express a gene, silence a gene, or simultaneously silence a first gene while expressing a second gene sequence. Please observe FIG.l, this first screen is preferred but is optional. The user may select one of these options and then be directed to another screen in which he or she is

able to insert nucleic acid information. If the user selects the option, silence the expression of a gene sequence, a screen appears with tabs labeled, "Gene Silencing RNA," "Gene Silencing Promoter," "Reporter Selection Gene," "Other Elements," "Envelope Pseudotype," and "Review Vector" as illustrated in FIG. 10. If the user selects the option, express a gene sequence, a screen appears with tabs labeled, "Gene," "Gene Expression Promoter," "Reporter/Selection Gene," "Other Elements," "Envelope Pseudotype," and "Review Vector." as illustrated in FIG. 2. If the user selects the option, simultaneously silence a first gene while expressing a second gene sequence, a screen appears with tabs labeled, "Gene Silencing RNA," "Gene Silencing Promoter," "Gene," "Gene Expressing Promoter," "Reporter/Selection Gene," "Other Elements," "Envelope Psuedotype," "Checkout." This screen is not illustrated. In each screen, it is preferable that all of these tabs are present; however, a graphic display could be constructed containing one or more of these tabs, or tabs in addition to the ones that are listed.

Gene Custom Lentiviral Vectors Each tab allows the user to insert a nucleic acid sequence into an area on the screen or graphic display. The nucleic acid sequence is preferable a biomolecule. As shown in FIG. 2, the first tab labeled, "Gene" is selected and an area is displayed for the insertion of a nucleic acid sequence of a biomolecule, preferably a gene. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule such as an "kinase", a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no." As shown in FIG. 2, there is a second tab labeled, "Gene Expressing Promoter" that may be selected to display an area, screen, for the insertion of a nucleic acid sequence of a biomolecule, preferably a promoter sequence. FIG. 3 illustrates one example of a screen that is displayed when the tab labeled "Gene Expressing Promoter" is selected by a user. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule, a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. As shown in FIG. 3, a user may click on one of three promoter names listed on the screen, that is, simian CMV (SCMV), Tet-off (Teto), or EF- lα (EF- lα) and the nucleic acid sequence of one of the

tnese biomolecules will be inserted into the space underneath the term "sequence." There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no."

As shown in FIG. 3, there is a third tab labeled, "Reporter/Selection Gene" that may be selected to display an area for the insertion of a nucleic acid sequence of a biomolecule, preferably either a reporter gene and/or a selection gene such as turbo-GFP (GFP). FIG. 4 illustrates one example of a screen, displayed when the tab labeled, "Reporter/Selection Gene" is selected by a user. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule such as a reporter gene or selection gene, a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no." As shown in FIG. 4, there is a fourth tab labeled, "Other Elements" that may be selected to display an area for the insertion of a nucleic acid sequence of a biomolecule, preferably an enhancer element such as a wood chuck element. FIG. 5 illustrates one example of an area displayed when the tab labeled, "Other Element" is selected by a user. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule, a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. As shown in FIG. 5, a user may click on the terms WPRE (WPRE) or truncated U3 sequence (SIN) to automatically insert the nucleic acid sequences of one of these biomolecules in the space located immediately under the term, "sequence." There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no."

As shown in FIG. 5, there is a fifth tab labeled, "Envelope Pseudotype" that may be selected to display an area for the insertion of a nucleic acid sequence of a biomolecule, preferably an envelope protein gene such as VSV-G (VSV) or Mokola-G (MOK). FIG. 6 illustrates one example of an area displayed when the tab labeled, "Envelope Pseutotype" is selected by a user. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule, a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth

space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. As shown in FIG. 6, a user may click on the terms VSV-G (VSV) or Mokola-G (MOK) to automatically insert the nucleic acid sequences of one of these biomolecules in the space located immediately under the term, "sequence." There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no."

As shown in FIG. 6, there is a sixth tab labeled, "Review Vector" that may be selected to display an area summarizing the design of the vector and corresponding pricing information related to the construction of this vector. FIG. 7 illustrates one example of an area displayed when the tab labeled, "Review Vector" is selected by a user. The user will observe a table having the headings, "Element," "Type," "Name," "Source," and "Costs." Under the heading labeled, "Element" is a list of biomolecules the user may have inserted into his or her custom vector. Under the heading labeled, "Type" is a list of additional information about the one or more biomolecule(s) selected by a user. Under the heading labeled, "Name" is a list of the names of the one or more biomolecule(s). Under the heading labeled, "source" is a list. Under the heading labeled, "Costs" is the price corresponding to each biomolecule inserted into the vector and the cost of the vector itself. A user click on the term "save" on the screen and safe the vector construct on the website or select to move to a Checkout screen.

FIGS. 8 and 9 illustrate one example of a Checkout screen of the present invention. The Checkout screen may include spaces for the user to provide shipping information, billing information, a promotional code obtained from a company representative or promotional item, purchase information (i.e. method of payment such as by credit card or purchase order), and shipping date information. In addition, the Checkout screen may include a "click agreement" that describes the terms and conditions of that apply when a custom vector is purchased off the web. As shown in FIG. 8, the click agreement is provided in a space under the terms, "Terms & Conditions." The Checkout screen may include a space a user may select to finalize an order. As shown in FIG. 8, a bar with the words, "Finalize Order" may be selected by a user.

RNAi Lentiviral Constructs Each tab allows the user to insert a nucleic acid sequence into an area on the screen or graphic display. The nucleic acid sequence is preferable a biomolecule. As shown in FIG. 10, the first tab labeled, "GeneSilencing RNA" is selected and an area is displayed for the insertion of a nucleic acid sequence of a biomolecule, preferably a gene silencing RNA. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of

biomolecule such as an "shRNA", a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no."

As shown in FIG. 10, there is a second tab labeled, "Gene Silencing Promoter" that may be selected to display an area for the insertion of a nucleic acid sequence of a biomolecule, preferably a promoter sequence. FIG. 11 illustrates one example of an area displayed when the tab labeled, "Gene Silencing Promoter" is selected by a user. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule such as Human Hl (Hl) and Human U6 (U6). promoter, a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no."

As shown in FIG. 11, there is a third tab labeled, "Reporter/Selection Gene" that may be selected to display an area for the insertion of a nucleic acid sequence of a biomolecule, preferably either a reporter gene and/or a selection gene such as turbo-GFP (GFP). FIG. 12 illustrates one example of an area displayed when the tab labeled, "Reporter/Selection Gene" is selected by a user. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule such as a reporter gene or selection gene, a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no."

As shown in FIG. 12 there is a fourth tab labeled, "Reporter Selection Gene Promoter" that may be selected to display an area for the insertion of a nucleic acid sequence of a biomolecule, preferably a gene promoter. FIG. 13 illustrates one example of an area displayed when the tab labeled, "Reporter Selection Gene Promoter" is selected by a user.

As shown in FIG. 13, there is a fourth tab labeled, "Other Elements" that may be selected to display an area for the insertion of a nucleic acid sequence of a biomolecule, preferably an enhancer element such as post-transcriptional elements and/or sequences to be included in the U3 region of the 3'LTR. FIG. 14 illustrates one example of an area displayed when the tab labeled,

"Other Elements" is selected by a user. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule, a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no."

As shown in FIG. 14, there is a fifth tab labeled, "Envelope Pseudotype" that may be selected to display an area for the insertion of a nucleic acid sequence of a biomolecule, preferably an envelope protein gene such as VSV-G (VSV) or Mokola-G (MOK). FIG. 15 illustrates one example of an area displayed when the tab labeled, "Envelope Psuedotype" is selected by a user. This area may include a first space for the name of a biomolecule, a second space for describing the "type" of biomolecule, a third space for allowing the user to "select" a biomolecule nucleic acid sequence that is already present on the graphic display, and a fourth space labeled "sequence" allowing the user to insert the nucleic acid sequence of the biomolecule named in the first space. There could also be a fifth space on the bottom of this screen that asks the question, "Will you provide DNA?" to which the user may either select "yes" or "no."

As shown in FIG. 15, there is a sixth tab labeled, "Review Vector" that may be selected to display an area summarizing the design of the vector and corresponding pricing information related to the construction of this vector. FIG. 16 illustrates one example of an area displayed when the tab labeled, "Review Vector" is selected by a user. The user will observe a table having the headings, "Element," "Type," "Name," "Source," and "Costs." Under the heading labeled, "Element" is a list of biomolecules the user may have inserted into his or her custom vector. Under the heading labeled, "Type" is a list of additional information about the one or more biomolecule(s) selected by a user. Under the heading labeled, "Name" is a list of the names of the one or more biomolecule(s). Under the heading labeled, "source" is a list. Under the heading labeled, "Costs" is the price corresponding to each biomolecule inserted into the vector and the cost of the vector itself. A user may click on the term "save" on the screen to save the vector construct on the website or select to move to a Checkout screen. Examples of such screens are illustrated in FIGS. 8 and 9.

In addition, web links may be included on any of the screens enabling a user, via the web, to connect directly to databases, preferably containing nucleic acid and protein sequences. The user could easily copy sequences from these databases and then paste them into spaces located on different screens, or areas, described above.

Although the present invention has been described in detail with reference to examples above, it is understood that various modifications can be made without departing from the spirit of the invention. Accordingly, the invention is limited only by the following claims. AU cited patents, patent applications and publications referred to in this application are herein expressly incorporated by reference in their entirety.